Knock-out of a mitochondrial sirtuin protects neurons from degeneration in Caenorhabditis elegans.

PubMed

Sangaletti, Rachele; D'Amico, Massimo; Grant, Jeff; Della-Morte, David; Bianchi, Laura

2017-08-01

Sirtuins are NAD⁺-dependent deacetylases, lipoamidases, and ADP-ribosyltransferases that link cellular metabolism to multiple intracellular pathways that influence processes as diverse as cell survival, longevity, and cancer growth. Sirtuins influence the extent of neuronal death in stroke. However, different sirtuins appear to have opposite roles in neuronal protection. In Caenorhabditis elegans, we found that knock-out of mitochondrial sirtuin sir-2.3, homologous to mammalian SIRT4, is protective in both chemical ischemia and hyperactive channel induced necrosis. Furthermore, the protective effect of sir-2.3 knock-out is enhanced by block of glycolysis and eliminated by a null mutation in daf-16/FOXO transcription factor, supporting the involvement of the insulin/IGF pathway. However, data in Caenorhabditis elegans cell culture suggest that the effects of sir-2.3 knock-out act downstream of the DAF-2/IGF-1 receptor. Analysis of ROS in sir-2.3 knock-out reveals that ROS become elevated in this mutant under ischemic conditions in dietary deprivation (DD), but to a lesser extent than in wild type, suggesting more robust activation of a ROS scavenging system in this mutant in the absence of food. This work suggests a deleterious role of SIRT4 during ischemic processes in mammals that must be further investigated and reveals a novel pathway that can be targeted for the design of therapies aimed at protecting neurons from death in ischemic conditions.

Genetic Contributors to Intergenerational CAG Repeat Instability in Huntington's Disease Knock-In Mice.

PubMed

Neto, João Luís; Lee, Jong-Min; Afridi, Ali; Gillis, Tammy; Guide, Jolene R; Dempsey, Stephani; Lager, Brenda; Alonso, Isabel; Wheeler, Vanessa C; Pinto, Ricardo Mouro

2017-02-01

Huntington's disease (HD) is a neurodegenerative disorder caused by the expansion of a CAG trinucleotide repeat in exon 1 of the HTT gene. Longer repeat sizes are associated with increased disease penetrance and earlier ages of onset. Intergenerationally unstable transmissions are common in HD families, partly underlying the genetic anticipation seen in this disorder. HD CAG knock-in mouse models also exhibit a propensity for intergenerational repeat size changes. In this work, we examine intergenerational instability of the CAG repeat in over 20,000 transmissions in the largest HD knock-in mouse model breeding datasets reported to date. We confirmed previous observations that parental sex drives the relative ratio of expansions and contractions. The large datasets further allowed us to distinguish effects of paternal CAG repeat length on the magnitude and frequency of expansions and contractions, as well as the identification of large repeat size jumps in the knock-in models. Distinct degrees of intergenerational instability were observed between knock-in mice of six background strains, indicating the occurrence of trans-acting genetic modifiers. We also found that lines harboring a neomycin resistance cassette upstream of Htt showed reduced expansion frequency, indicative of a contributing role for sequences in cis, with the expanded repeat as modifiers of intergenerational instability. These results provide a basis for further understanding of the mechanisms underlying intergenerational repeat instability. Copyright © 2017 by the Genetics Society of America.

[Role of the sweet taste receptor in glucose metabolism: no sweets for diabetes?].

PubMed

Nomura, Masatoshi; Kawahara, Yuta

2015-01-01

Type 2 diabetes is closely associated with our daily diets and has become a global health problem with increasing number of patients. Maintaining energy homeostasis is essentially required for the treatment of diabetes. Energy metabolism starts with taking in a meal. Nutrients including amino acids, fatty acids and glucose in the digest have been shown to act on the neuroendocrine cells in the gastrointestinal (GI) tract, and thereby play important roles in energy homeostasis. Therefore, the GI tract is now recognized as a sensor system for nutrient signals. Taste receptor type 1 member 2 (T1R2) is known to function as a co-receptor with T1R3 to detect sweet chemicals in the taste buds. It has been proposed that the T1R2/T1R3 receptor complex acts as sweet sensor in the intestine, and plays a pivotal role in sensing sugars and maintaining glucose homeostasis through incretin secretion. To clarify the physiological roles of T1R2 in glucose homeostasis, T1r2-lacZ knock-in/knock-out mice were generated. We found lacZ gene expression in the GI tract where T1r3 expression has been reported. Interestingly, the T1r2-lacZ knock-in mice showed impaired glucose tolerance on oral glucose challenge but not on intraperitoneal injection. However, the fasting glucose level in T1r2-lacZ knock-in mice was comparable to that in wild type mice. These results suggest an important role of the sweet taste receptor system in the intestine when stimulated by glucose. Therefore, the roles of T1R2 will be presented and the mechanism for metabolic homeostasis will be discussed.

Opportunity Knocks: Pipeline Programs Offer Minority Students a Path to Dentistry

ERIC Educational Resources Information Center

Fauteux, Nicole

2012-01-01

Minority students have traditionally been underrepresented in dental schools, which is why enrichment and pipeline programs aimed at helping minority students are necessary. That reality is reflected in their woeful underrepresentation among practicing dentists. Hispanics made up only 5.8 percent of practicing dentists in 2011, according to the…

What Not to Do during a Presidential Interview

ERIC Educational Resources Information Center

Pulliams, Preston

2016-01-01

As a presidential search consultant, Preston Pulliams has had the opportunity to observe and participate in many presidential search interviews and the meetings where hiring decisions are made. He has observed how some interview candidates simply knocked themselves out of contention by employing one or more of five poor interview strategies. In…

A Presidential Search Is Opportunity Knocking

ERIC Educational Resources Information Center

Funk, R. William

2005-01-01

When the inevitable change in presidential leadership takes place at a college or university, board members often think only of the additional demands the search will place on their hectic schedules. No other situation allows board members to contribute so positively and profoundly to the immediate and long-term welfare of the college or…

Knock-out of a mitochondrial sirtuin protects neurons from degeneration in Caenorhabditis elegans

PubMed Central

Sangaletti, Rachele; Grant, Jeff; Della-Morte, David

2017-01-01

Sirtuins are NAD⁺-dependent deacetylases, lipoamidases, and ADP-ribosyltransferases that link cellular metabolism to multiple intracellular pathways that influence processes as diverse as cell survival, longevity, and cancer growth. Sirtuins influence the extent of neuronal death in stroke. However, different sirtuins appear to have opposite roles in neuronal protection. In Caenorhabditis elegans, we found that knock-out of mitochondrial sirtuin sir-2.3, homologous to mammalian SIRT4, is protective in both chemical ischemia and hyperactive channel induced necrosis. Furthermore, the protective effect of sir-2.3 knock-out is enhanced by block of glycolysis and eliminated by a null mutation in daf-16/FOXO transcription factor, supporting the involvement of the insulin/IGF pathway. However, data in Caenorhabditis elegans cell culture suggest that the effects of sir-2.3 knock-out act downstream of the DAF-2/IGF-1 receptor. Analysis of ROS in sir-2.3 knock-out reveals that ROS become elevated in this mutant under ischemic conditions in dietary deprivation (DD), but to a lesser extent than in wild type, suggesting more robust activation of a ROS scavenging system in this mutant in the absence of food. This work suggests a deleterious role of SIRT4 during ischemic processes in mammals that must be further investigated and reveals a novel pathway that can be targeted for the design of therapies aimed at protecting neurons from death in ischemic conditions. PMID:28820880

Is Opportunity Knocking or Slipping Away? Racial Diversity and Segregation in Pennsylvania

ERIC Educational Resources Information Center

Kotok, Stephen; Reed, Katherine

2015-01-01

Historically, Pennsylvania has struggled to integrate its public schools, especially with much of the racial diversity concentrated in urban regions. Starting in the 1960s, the Pennsylvania Human Relations Commission (PHRC) was the state's enforcing body to combat school desegregation, but since the early 1980s, when it comes to education, the…

Opportunity is Knocking: Will Education Open the Door? Carnegie Perspectives

ERIC Educational Resources Information Center

Iiyoshi, Toru

2006-01-01

This essay presents a discussion of how the tools and resources of open source education may demonstrably improve education quality. The main tenet of open education is to make educational assets freely available to the public. This is becoming easier and less expensive as network and multimedia technology evolves. Obstacles may stand in the way…

Opportunity Knocks! Inquiry, the New National Social Studies and Science Standards, and You

ERIC Educational Resources Information Center

Ratzer, Mary Boyd

2014-01-01

To be recognized and adopted, the promise and potential of national standards must validate the instructional role of the school librarian and build formative knowledge through authentic process and products. Inquiry is a direct link to those dimensions. Standards that are rigorous and relevant optimize the developing brain's affinity for building…

When Opportunity Knocks Twice: Two-Year-Olds' Repetition of Sentence Subjects

ERIC Educational Resources Information Center

Valian, Virginia; Aubry, Stephanie

2005-01-01

Why are young children's utterances short? This elicited imitation study used a new task--double imitation--to investigate the factors that contribute to children's failure to lexicalize sentence subjects. Two-year-olds heard a triad of sentences singly and attempted to imitate each; they then again heard the same triad singly and again attempted…

Opportunity Knocks? The Impact of Bursary Schemes on Students from Low-Income Backgrounds

ERIC Educational Resources Information Center

Hatt, Sue; Hannan, Andrew; Baxter, Arthur; Harrison, Neil

2005-01-01

In England, the government target that 50% of young people should gain experience of higher education has prompted many initiatives to widen participation. National policies, however, are often implemented at institutional or local level. As a result, the impact upon the individual participants can vary according to the context in which the…

When a Leadership Opportunity Knocks, Answer!

ERIC Educational Resources Information Center

DelGuidice, Margaux

2011-01-01

In the fall of 2010 the author received an e-mail informing her that she has been selected as a Gale/Cengage New Leader, which was a true honor. Not only would she be joining four dynamic school librarians from around the country at the School Library Journal Summit on the Future of Reading, she was also being paired with a mentor to navigate her…

When Opportunity Knocks, Who Answers? New Evidence on College Achievement Awards. NBER Working Paper No. 16643

ERIC Educational Resources Information Center

Angrist, Joshua; Oreopoulos, Philip; Williams, Tyler

2010-01-01

We evaluate the effects of academic achievement awards for first and second-year college students on a Canadian commuter campus. The award scheme offered linear cash incentives for course grades above 70. Awards were paid every term. Program participants also had access to peer advising by upperclassmen. Program engagement appears to have been…

Knocking on the Doors of Opportunity: A Phenomenological Study of How African American Males Have Experienced Their Journey to the Community College Presidency

ERIC Educational Resources Information Center

Bugg, Elmer A., Jr.

2009-01-01

This qualitative study elicited key themes in an attempt to understand the nature and essence of the career advancement experiences of a select group of African American male community college presidents. Research was conducted through a "culturally sensitive" research design (Tillman, 2002) that contained four elements: "culturally congruent…

Respiratory Home Health Care

MedlinePlus

... if it gets knocked over and damaged, the gas could escape, making the cylinder act like a rocket. If you have long tubing that lets you move about the house, warn visitors so they won’t trip on it. If you have a vessel of liquid oxygen, keep it upright. If it falls on ...

Suppression of the hypothalamic-pituitary-gonadal axis by TAK-385 (relugolix), a novel, investigational, orally active, small molecule gonadotropin-releasing hormone (GnRH) antagonist: studies in human GnRH receptor knock-in mice.

PubMed

Nakata, Daisuke; Masaki, Tsuneo; Tanaka, Akira; Yoshimatsu, Mie; Akinaga, Yumiko; Asada, Mari; Sasada, Reiko; Takeyama, Michiyasu; Miwa, Kazuhiro; Watanabe, Tatsuya; Kusaka, Masami

2014-01-15

TAK-385 (relugolix) is a novel, non-peptide, orally active gonadotropin-releasing hormone (GnRH) antagonist, which builds on previous work with non-peptide GnRH antagonist TAK-013. TAK-385 possesses higher affinity and more potent antagonistic activity for human and monkey GnRH receptors compared with TAK-013. Both TAK-385 and TAK-013 have low affinity for the rat GnRH receptor, making them difficult to evaluate in rodent models. Here we report the human GnRH receptor knock-in mouse as a humanized model to investigate pharmacological properties of these compounds on gonadal function. Twice-daily oral administration of TAK-013 (10mg/kg) for 4 weeks decreased the weights of testes and ventral prostate in male knock-in mice but not in male wild-type mice, demonstrating the validity of this model to evaluate antagonists for the human GnRH receptor. The same dose of TAK-385 also reduced the prostate weight to castrate levels in male knock-in mice. In female knock-in mice, twice-daily oral administration of TAK-385 (100mg/kg) induced constant diestrous phases within the first week, decreased the uterus weight to ovariectomized levels and downregulated GnRH receptor mRNA in the pituitary after 4 weeks. Gonadal function of TAK-385-treated knock-in mice began to recover after 5 days and almost completely recovered within 14 days after drug withdrawal in both sexes. Our findings demonstrate that TAK-385 acts as an antagonist for human GnRH receptor in vivo and daily oral administration potently, continuously and reversibly suppresses the hypothalamic-pituitary-gonadal axis. TAK-385 may provide useful therapeutic interventions in hormone-dependent diseases including endometriosis, uterine fibroids and prostate cancer. Copyright © 2013 Elsevier B.V. All rights reserved.

Knock-Down of Cathepsin D Affects the Retinal Pigment Epithelium, Impairs Swim-Bladder Ontogenesis and Causes Premature Death in Zebrafish

PubMed Central

Follo, Carlo; Ozzano, Matteo; Mugoni, Vera; Castino, Roberta; Santoro, Massimo; Isidoro, Ciro

2011-01-01

The lysosomal aspartic protease Cathepsin D (CD) is ubiquitously expressed in eukaryotic organisms. CD activity is essential to accomplish the acid-dependent extensive or partial proteolysis of protein substrates within endosomal and lysosomal compartments therein delivered via endocytosis, phagocytosis or autophagocytosis. CD may also act at physiological pH on small-size substrates in the cytosol and in the extracellular milieu. Mouse and fruit fly CD knock-out models have highlighted the multi-pathophysiological roles of CD in tissue homeostasis and organ development. Here we report the first phenotypic description of the lack of CD expression during zebrafish (Danio rerio) development obtained by morpholino-mediated knock-down of CD mRNA. Since the un-fertilized eggs were shown to be supplied with maternal CD mRNA, only a morpholino targeting a sequence containing the starting ATG codon was effective. The main phenotypic alterations produced by CD knock-down in zebrafish were: 1. abnormal development of the eye and of retinal pigment epithelium; 2. absence of the swim-bladder; 3. skin hyper-pigmentation; 4. reduced growth and premature death. Rescue experiments confirmed the involvement of CD in the developmental processes leading to these phenotypic alterations. Our findings add to the list of CD functions in organ development and patho-physiology in vertebrates. PMID:21747967

When Opportunity Only Knocks Once: Lessons From Gen Petraeus In Iraq And Afghanistan

DTIC Science & Technology

2016-03-01

Afghanistan because of the unfavorable contextual elements including the lack of both popular and political support at home and abroad, the tribal...decades-long, war ravaged land in Afghanistan. Different contextual environments and battlefield elements can stymie a war effort of an otherwise...37th Commandant of the U.S.M.C. GEN David Petraeus’ counterinsurgency (COIN) efforts failed in Afghanistan because of the unfavorable contextual

Zika virus infection of adult and fetal STAT2 knock-out hamsters.

PubMed

Siddharthan, Venkatraman; Van Wettere, Arnaud J; Li, Rong; Miao, Jinxin; Wang, Zhongde; Morrey, John D; Julander, Justin G

2017-07-01

Zika virus (ZIKV) infection was investigated in adult and fetal STAT2 knock-out (KO) hamsters. Subcutaneous injection of ZIKV of adults resulted in morbidity, mortality, and infection of the uterus, placenta, brain, spinal cord, and testicles, thus providing an opportunity to evaluate congenital ZIKV infection in a second rodent species besides mice. ZIKV-infected cells with morphologies of Sertoli cells and spermatogonia were observed in the testes, which may have implications for sexual transmission and male sterility. Neonates exposed as fetuses to ZIKV at 8 days post-coitus were not smaller than controls. Nevertheless, infectious virus and ZIKV RNA was detected in some, but not all, placentas and fetal brains of KO hamsters. STAT2 KO hamsters may be useful for addressing sexual transmission, pathogenesis, routes of fetal infection, and neurological disease outcomes, and may also be used in antiviral or vaccine studies to identify intervention strategies. Copyright © 2017. Published by Elsevier Inc.

Knock-down and speed of kill of a combination of fipronil and permethrin for the prevention of Ctenocephalides felis flea infestation in dogs.

PubMed

Halos, Lénaïg; Fourie, Josephus J; Fankhauser, Becky; Beugnet, Frederic

2016-02-02

A topical combination of fipronil + permethrin (Frontline Tri-Act/Frontect, Merial) has recently been developed to control fleas, ticks, mosquitoes, sandflies and stable flies on dogs. Two studies were conducted to assess its speed of kill and knock-down effect on Ctenocephalides felis fleas. The combination was compared to either fipronil alone or to a combination of permethrin, dinotefuran, and pyriproxyfen, In each study, 18 dogs were randomly allocated to one of three groups: (Group 1: untreated dog; Group 2: treated once on D0 with the combination of fipronil and permethrin; Group 3: treated once on D0 either with fipronil alone (study 1) or with a combination of permethrin, dinetofuran and pyriproxyfen (study 2)). Each dog was infested with 100 unfed adult C. felis fleas on Days 2 (study 2), 7, 14, 21 and 28. Fleas were collected from dogs at 1 h and 12 h post- infestations (PI) (study 1) or at 2 h and 6 h PI (study 2) to assess efficacy and from collection pans underneath cages 1 h (study 1) or 5 min (study 2) PI to assess knock-down effect. All treated dogs had significantly (p ≤ 0.01) lower flea counts than untreated dogs at every time point in both studies. For a whole month, a significant knock-down effect against infesting fleas is obtained in five minutes PI with the combination of permethrin and fipronil. Complete efficacy (>95%) was achieved in 1 h (study 1) or 2 h (study 2) PI for 14 days and by 6 h PI for all challenges conducted throughout the month. Efficacy remains >85% at 2 h PI for the whole month. A significantly higher efficacy of the fipronil + permethrin combination compared to other treatments was demonstrated at the earliest time points for the month (1 h knock-down effect and insecticidal efficacy compared to fipronil alone; 5 min knock-down effect compared to the combination of permethrin + dinetofuran + pyriproxyfen). The rapid flea knock-down effect and speed of kill demonstrated by the spot on combination of fipronil + permethrin provide a reliable strategy against flea infestations in dogs.

Opportunity Knocks: Closing the Gaps between Leaders and the Public on Math, Science, & Technology Education--A Qualitative Research Report on the Kansas City Region

ERIC Educational Resources Information Center

Kadlec, Alison; Friedman, Will

2010-01-01

The Kansas City region is in many ways representative of the larger national economy. As with many cities of its size, the largest area of employment, other than in government, is in the healthcare sector. While biomedical research is a major growth area across the nation, Kansas City is also the national leader in the animal healthcare industry,…

76 FR 67033 - To Take Certain Actions Under the African Growth and Opportunity Act

Federal Register 2010, 2011, 2012, 2013, 2014

2011-10-28

... Actions Under the African Growth and Opportunity Act #0; #0; #0; Presidential Documents #0; #0; #0;#0... Under the African Growth and Opportunity Act By the President of the United States of America A...)(1)), as added by section 111(a) of the African Growth and Opportunity Act (title I of Public Law 106...

The School-to-Work Opportunities Act: An Opportunity To Serve All Students. Women's Educational Equity Act Publishing Center Digest.

ERIC Educational Resources Information Center

Wiberg, Mary

This digest consists of two brief articles. The lead article, "The School-to-Work Opportunities Act: An Opportunity To Serve All Students" (Mary Wiberg) tells how the STW Act differs from vocational education by expanding the traditional programs and reflecting the constructivist model of education reform and how STW meets the needs of…

The ergonomic conundrum for technology integration.

PubMed

Ahearn, David

2006-01-01

In the operatory of today, the opportunities are great. Space is at a premium and integration is a must. By consolidating the operator's primary supplies, which formerly were spread around the room, performance improves and space is gained for the successful introduction of high technology. Room cost decreases, permitting more networked rooms in less space at a lower cost. The productive potential of today's practice is greater than ever before; however, to gain the most from these opportunities, a change must occur in our perception of the treatment environment. When designing an office for the coming decades, it is vitally important to create a space for the future, not the past. Dentists must be willing to look objectively at their clinical environment and be willing to gain a clear sense of future needs. Your new treatment room can become a tool for future growth and prosperity or a museum about dentistry's past. Opportunity knocks.

75 FR 32480 - Funding Opportunity: Affordable Care Act Medicare Beneficiary Outreach and Assistance Program...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-06-08

... Care Act Medicare Beneficiary Outreach and Assistance Program Funding for Title VI Native American Programs Purpose of Notice: Availability of funding opportunity announcement. Funding Opportunity Title/Program Name: Affordable Care Act Medicare Beneficiary Outreach and Assistance Program Funding for Title...

CCN3 Protein Participates in Bone Regeneration as an Inhibitory Factor*

PubMed Central

Matsushita, Yuki; Sakamoto, Kei; Tamamura, Yoshihiro; Shibata, Yasuaki; Minamizato, Tokutaro; Kihara, Tasuku; Ito, Masako; Katsube, Ken-ichi; Hiraoka, Shuichi; Koseki, Haruhiko; Harada, Kiyoshi; Yamaguchi, Akira

2013-01-01

CCN3, a member of the CCN protein family, inhibits osteoblast differentiation in vitro. However, the role of CCN3 in bone regeneration has not been well elucidated. In this study, we investigated the role of CCN3 in bone regeneration. We identified the Ccn3 gene by microarray analysis as a highly expressed gene at the early phase of bone regeneration in a mouse bone regeneration model. We confirmed the up-regulation of Ccn3 at the early phase of bone regeneration by RT-PCR, Western blot, and immunofluorescence analyses. Ccn3 transgenic mice, in which Ccn3 expression was driven by 2.3-kb Col1a1 promoter, showed osteopenia compared with wild-type mice, but Ccn3 knock-out mice showed no skeletal changes compared with wild-type mice. We analyzed the bone regeneration process in Ccn3 transgenic mice and Ccn3 knock-out mice by microcomputed tomography and histological analyses. Bone regeneration in Ccn3 knock-out mice was accelerated compared with that in wild-type mice. The mRNA expression levels of osteoblast-related genes (Runx2, Sp7, Col1a1, Alpl, and Bglap) in Ccn3 knock-out mice were up-regulated earlier than those in wild-type mice, as demonstrated by RT-PCR. Bone regeneration in Ccn3 transgenic mice showed no significant changes compared with that in wild-type mice. Phosphorylation of Smad1/5 was highly up-regulated at bone regeneration sites in Ccn3 KO mice compared with wild-type mice. These results indicate that CCN3 is up-regulated in the early phase of bone regeneration and acts as a negative regulator for bone regeneration. This study may contribute to the development of new strategies for bone regeneration therapy. PMID:23653360

DOE Office of Scientific and Technical Information (OSTI.GOV)

Pamminger, Michael; Sevik, James; Scarcelli, Riccardo

The compression ratio is a strong lever to increase the efficiency of an internal combustion engine. However, among others, it is limited by the knock resistance of the fuel used. Natural gas shows a higher knock resistance compared to gasoline, which makes it very attractive for use in internal combustion engines. The current paper describes the knock behavior of two gasoline fuels, and specific incylinder blend ratios with one of the gasoline fuels and natural gas. The engine used for these investigations is a single cylinder research engine for light duty application which is equipped with two separate fuel systems.more » Both fuels can be used simultaneously which allows for gasoline to be injected into the intake port and natural gas to be injected directly into the cylinder to overcome the power density loss usually connected with port fuel injection of natural gas. Adding natural gas at wide open throttle helps to reduce knock mitigating measures and increases the efficiency and power density compared to the other gasoline type fuels with lower knock resistance. The used methods, knock intensity and number of pressure waves, do not show significant differences in knock behavior for the natural gas - gasoline blends compared to the gasoline type fuels. A knock integral was used to describe the knock onset location of the fuels tested. Two different approaches were used to determine the experimental knock onset and were compared to the knock onset delivered by the knock integral (chemical knock onset). The gasoline type fuels show good agreement between chemical and experimental knock onset. However, the natural gas -gasoline blends show higher discrepancies comparing chemical and experimental knock onset.« less

Functional genetics for all: engineered nucleases, CRISPR and the gene editing revolution.

PubMed

Gilles, Anna F; Averof, Michalis

2014-01-01

Developmental biology, as all experimental science, is empowered by technological advances. The availability of genetic tools in some species - designated as model organisms - has driven their use as major platforms for understanding development, physiology and behavior. Extending these tools to a wider range of species determines whether (and how) we can experimentally approach developmental diversity and evolution. During the last two decades, comparative developmental biology (evo-devo) was marked by the introduction of gene knockdown and deep sequencing technologies that are applicable to a wide range of species. These approaches allowed us to test the developmental role of specific genes in diverse species, to study biological processes that are not accessible in established models and, in some cases, to conduct genome-wide screens that overcome the limitations of the candidate gene approach. The recent discovery of CRISPR/Cas as a means of precise alterations into the genome promises to revolutionize developmental genetics. In this review we describe the development of gene editing tools, from zinc-finger nucleases to TALENs and CRISPR, and examine their application in gene targeting, their limitations and the opportunities they present for evo-devo. We outline their use in gene knock-out and knock-in approaches, and in manipulating gene functions by directing molecular effectors to specific sites in the genome. The ease-of-use and efficiency of CRISPR in diverse species provide an opportunity to close the technology gap that exists between established model organisms and emerging genetically-tractable species.

7 CFR 4279.58 - Equal Credit Opportunity Act.

Code of Federal Regulations, 2010 CFR

2010-01-01

... (providing the applicant has the capacity to contract), or because all or part of the applicant's income derives from a public assistance program, or because the applicant has, in good faith, exercised any right... Opportunity Act. In accordance with title V of Public Law 93-495, the Equal Credit Opportunity Act, with...

78 FR 26650 - Agency Information Collection Activities: African Growth and Opportunity Act Certificate of Origin

Federal Register 2010, 2011, 2012, 2013, 2014

2013-05-07

... Activities: African Growth and Opportunity Act Certificate of Origin AGENCY: U.S. Customs and Border... and Opportunity Act Certificate of Origin (AGOA). This request for comments is being made pursuant to... submitted will be summarized and included in the CBP request for Office of Management and Budget (OMB...

Environmental Threats to Security, Stability, and U.S. Interests in Southern Africa: Opportunity Knocks - Time for a Comprehensive Region Defense Environmental International Cooperation and Environmental Security Assistance Strategy

DTIC Science & Technology

2009-01-01

Cattle raiding and the effects of poaching would likely intensify as well. Fisheries may be adversely impacted due to climatic effects on coral...organized crime to set roots in southern Africa during the early 1990s. Chinese gangs have since diversified into trafficking of poached rhino horn...charged with poaching rhino horn for shipment to China.90 Intelligence sources developed by Botswana security personnel for anti- poaching purposes

The Impact of Low Octane Hydrocarbon Blending Streams on Ethanol Engine Optimization

DOE Office of Scientific and Technical Information (OSTI.GOV)

Szybist, James P; West, Brian H

2013-01-01

Ethanol is a very attractive fuel from an end-use perspective because it has a high chemical octane number and a high latent heat of vaporization. When an engine is optimized to take advantage of these fuel properties, both efficiency and power can be increased through higher compression ratio, direct fuel injection, higher levels of boost, and a reduced need for enrichment to mitigate knock or protect the engine and aftertreatment system from overheating. The ASTM D5798 specification for high level ethanol blends, commonly called E85, underwent a major revision in 2011. The minimum ethanol content was revised downward from 68more » vol% to 51 vol%, which combined with the use of low octane blending streams such as natural gasoline introduces the possibility of a lower octane E85 fuel. While this fuel is suitable for current ethanol tolerant flex fuel vehicles, this study experimentally examines whether engines can still be aggressively optimized for the resultant fuel from the revised ASTM D5798 specification. The performance of six ethanol fuel blends, ranging from 51-85% ethanol, is compared to a premium-grade certification gasoline (UTG-96) in a single-cylinder direct-injection (DI) engine with a compression ratio of 12.9:1 at knock-prone engine conditions. UTG-96 (RON = 96.1), light straight run gasoline (RON = 63.6), and n-heptane (RON = 0) are used as the hydrocarbon blending streams for the ethanol-containing fuels in an effort to establish a broad range of knock resistance for high ethanol fuels. Results show that nearly all ethanol-containing fuels are more resistant to engine knock than UTG-96 (the only exception being the ethanol blend with 49% n-heptane). This knock resistance allows ethanol blends made with 33 and 49% light straight run gasoline, and 33% n-heptane to be operated at significantly more advanced combustion phasing for higher efficiency, as well as at higher engine loads. While experimental results show that the octane number of the hydrocarbon blend stock does impact engine performance, there remains a significant opportunity for engine optimization when considering even the lowest octane fuels that are in compliance with the current revision of ASTM D5798 compared to premium-grade gasoline.« less

Myosin Light Chain Kinase Is Necessary for Tonic Airway Smooth Muscle Contraction*

PubMed Central

Zhang, Wen-Cheng; Peng, Ya-Jing; Zhang, Gen-Sheng; He, Wei-Qi; Qiao, Yan-Ning; Dong, Ying-Ying; Gao, Yun-Qian; Chen, Chen; Zhang, Cheng-Hai; Li, Wen; Shen, Hua-Hao; Ning, Wen; Kamm, Kristine E.; Stull, James T.; Gao, Xiang; Zhu, Min-Sheng

2010-01-01

Different interacting signaling modules involving Ca2+/calmodulin-dependent myosin light chain kinase, Ca2+-independent regulatory light chain phosphorylation, myosin phosphatase inhibition, and actin filament-based proteins are proposed as specific cellular mechanisms involved in the regulation of smooth muscle contraction. However, the relative importance of specific modules is not well defined. By using tamoxifen-activated and smooth muscle-specific knock-out of myosin light chain kinase in mice, we analyzed its role in tonic airway smooth muscle contraction. Knock-out of the kinase in both tracheal and bronchial smooth muscle significantly reduced contraction and myosin phosphorylation responses to K+-depolarization and acetylcholine. Kinase-deficient mice lacked bronchial constrictions in normal and asthmatic airways, whereas the asthmatic inflammation response was not affected. These results indicate that myosin light chain kinase acts as a central participant in the contractile signaling module of tonic smooth muscle. Importantly, contractile airway smooth muscles are necessary for physiological and asthmatic airway resistance. PMID:20018858

Criminal Acts Against Civil Aviation. 1993

DTIC Science & Technology

1993-01-01

and the Canadian passenger. The hijacker was injured in the head , the pilot in the leg, and the passenger in the shoulder . It was later determined...er’s "accomplice" would destroy the aircraft unless the demand was met. The hijacker was carrying a shampoo bottle containing a mixture of hydrochloric...hijacker on the head with a whiskey bottle, knocking the man to the floor, and security officers took the hijacker from the aircraft. This was only the

The intensity of knock in an internal combustion engine: An experimental and modeling study

NASA Astrophysics Data System (ADS)

Cowart, J. S.; Haghooie, M.; Newman, C. E.; Davis, G. C.; Pitz, W. J.; Westbrook, C. K.

1992-09-01

Experimental data have been obtained that characterize knock occurrence times and knock intensities in a spark ignition engine operating on indolene and 91 primary reference fuel, as spark timing and inlet temperature were varied. Individual, in-cylinder pressure histories measured under knocking conditions were conditioned and averaged to obtain representative pressure traces. These averaged pressure histories were used as input to a reduced and detailed chemical kinetic model. The time derivative of CO concentration and temperature were correlated with the measured knock intensity and percent cycles knocking. The goal was to evaluate the potential of using homogeneous, chemical kinetic models as predictive tools for knock intensity.

The Personal Responsibility and Work Opportunity Reconciliation Act of 1996: An Examination of Its Impact on Legal Immigrants and Refugees in Rhode Island.

ERIC Educational Resources Information Center

Serpa, Fernando A.

This report describes a 1998 consultation conducted to examine the impact of the Personal Responsibility and Work Opportunity Reconciliation Act of 1996 on legal immigrants and refugees in Rhode Island. The Personal Responsibility and Work Opportunity Act restricted access of documented immigrants to a wide range of government programs such as…

76 FR 54774 - Notice of Intent To Award Affordable Care Act Funding, Funding Opportunity Announcement CDC-RFA...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-09-02

... Intent To Award Affordable Care Act Funding, Funding Opportunity Announcement CDC-RFA-DP10-1014 AGENCY... Funding Opportunity CDC-RFA- DP10-1014. It is the intent of CDC to provide continuation funding to one (1... published in the above referenced REACH CORE Funding Opportunity Announcement (FOA). Award Information...

12 CFR 268.101 - General policy for equal opportunity.

Code of Federal Regulations, 2010 CFR

2010-01-01

... Discrimination in Employment Act (ADEA) (29 U.S.C. 621 et seq.), the Equal Pay Act (29 U.S.C. 206(d)), or the... 12 Banks and Banking 3 2010-01-01 2010-01-01 false General policy for equal opportunity. 268.101... RESERVE SYSTEM RULES REGARDING EQUAL OPPORTUNITY Board Program To Promote Equal Opportunity § 268.101...

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sileghem, L.; Wallner, T.; Verhelst, S.

As knock is one of the main factors limiting the efficiency of spark-ignition engines, the introduction of alcohol blends could help to mitigate knock concerns due to the elevated knock resistance of these blends. A model that can accurately predict their autoignition behavior would be of great value to engine designers. The current work aims to develop such a model for alcohol–gasoline blends. First, a mixing rule for the autoignition delay time of alcohol–gasoline blends is proposed. Subsequently, this mixing rule is used together with an autoignition delay time correlation of gasoline and an autoignition delay time cor-relation of methanolmore » in a knock integral model that is implemented in a two-zone engine code. The pre-dictive performance of the resulting model is validated through comparison against experimental measurements on a CFR engine for a range of gasoline–methanol blends. The knock limited spark advance, the knock intensity, the knock onset crank angle and the value of the knock integral at the experimental knock onset have been simulated and compared to the experimental values derived from in-cylinder pressure measurements.« less

76 FR 164 - Agency Information Collection Activities: Haitian Hemispheric Opportunity Through Partnership...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-01-03

... (``Haiti HOPE'') Act of 2006. This is a proposed extension of an information collection that was previously... Encouragement (``Haiti HOPE'') Act of 2006. OMB Number: 1651-0129. Abstract: Title V of the Tax Relief and... Hemispheric Opportunity through Partnership Encouragement Act of 2006 (``Haiti HOPE Act''), provides for duty...

Optogenetic analysis of a nociceptor neuron and network reveals ion channels acting downstream of primary sensors.

PubMed

Husson, Steven J; Costa, Wagner Steuer; Wabnig, Sebastian; Stirman, Jeffrey N; Watson, Joseph D; Spencer, W Clay; Akerboom, Jasper; Looger, Loren L; Treinin, Millet; Miller, David M; Lu, Hang; Gottschalk, Alexander

2012-05-08

Nociception generally evokes rapid withdrawal behavior in order to protect the tissue from harmful insults. Most nociceptive neurons responding to mechanical insults display highly branched dendrites, an anatomy shared by Caenorhabditis elegans FLP and PVD neurons, which mediate harsh touch responses. Although several primary molecular nociceptive sensors have been characterized, less is known about modulation and amplification of noxious signals within nociceptor neurons. First, we analyzed the FLP/PVD network by optogenetics and studied integration of signals from these cells in downstream interneurons. Second, we investigated which genes modulate PVD function, based on prior single-neuron mRNA profiling of PVD. Selectively photoactivating PVD, FLP, and downstream interneurons via Channelrhodopsin-2 (ChR2) enabled the functional dissection of this nociceptive network, without interfering signals by other mechanoreceptors. Forward or reverse escape behaviors were determined by PVD and FLP, via integration by command interneurons. To identify mediators of PVD function, acting downstream of primary nocisensor molecules, we knocked down PVD-specific transcripts by RNAi and quantified light-evoked PVD-dependent behavior. Cell-specific disruption of synaptobrevin or voltage-gated Ca(2+) channels (VGCCs) showed that PVD signals chemically to command interneurons. Knocking down the DEG/ENaC channel ASIC-1 and the TRPM channel GTL-1 indicated that ASIC-1 may extend PVD's dynamic range and that GTL-1 may amplify its signals. These channels act cell autonomously in PVD, downstream of primary mechanosensory molecules. Our work implicates TRPM channels in modifying excitability of and DEG/ENaCs in potentiating signal output from a mechano-nociceptor neuron. ASIC-1 and GTL-1 homologs, if functionally conserved, may denote valid targets for novel analgesics. Copyright © 2012 Elsevier Ltd. All rights reserved.

78 FR 26031 - Sunshine Act Notice

Federal Register 2010, 2011, 2012, 2013, 2014

2013-05-03

... EQUAL EMPLOYMENT OPPORTUNITY COMMISSION Sunshine Act Notice AGENCY HOLDING THE MEETING: Equal Employment Opportunity Commission DATE AND TIME: Wednesday, May 8, 2013, 9:00 a.m. Eastern Time. PLACE.... Announcement of Notation Votes, and 2. Wellness Programs Under Federal Equal Employment Opportunity Laws. Note...

Implementation and Reauthorization of the Personal Responsibility and Work Opportunity Reconciliation Act of 1996. Hearing before the Committee on Indian Affairs on the Implementation and Reauthorization of the Personal Responsibility and Work Opportunity Reconciliation Act of 1996 in Indian Country. United States Senate, One Hundred Seventh Congress, Second Session.

ERIC Educational Resources Information Center

Congress of the U.S., Washington, DC. Senate Committee on Indian Affairs.

A Senate committee hearing received testimony on the implementation of the Personal Responsibility and Work Opportunity Reconciliation Act of 1996 in American Indian and Alaska Native tribal communities. The act provided authority for tribal governments to administer welfare programs formerly administered by the states but did not provide funds to…

76 FR 7562 - Sunshine Act Meeting

Federal Register 2010, 2011, 2012, 2013, 2014

2011-02-10

... EQUAL EMPLOYMENT OPPORTUNITY COMMISSION Sunshine Act Meeting AGENCY HOLDING THE MEETING: Equal Employment Opportunity Commission. DATE AND TIME: Wednesday, February 16, 2011, 9:30 a.m. Eastern Time. PLACE.... Announcement of Notation Votes, and 2. Out of work, out of luck? Denying employment opportunities to unemployed...

Knock detection system to improve petrol engine performance, using microphone sensor

NASA Astrophysics Data System (ADS)

Sujono, Agus; Santoso, Budi; Juwana, Wibawa Endra

2017-01-01

An increase of power and efficiency of spark ignition engines (petrol engines) are always faced with the problem of knock. Even the characteristics of the engine itself are always determined from the occurrence of knock. Until today, this knocking problem has not been solved completely. Knock is caused by principal factors that are influenced by the engine rotation, the load or opening the throttle and spark advance (ignition timing). In this research, the engine is mounted on the engine test bed (ETB) which is equipped with the necessary sensors. Knock detection using a new method, which is based on pattern recognition, which through the knock sound detection by using a microphone sensor, active filter, the regression of the normalized envelope function, and the calculation of the Euclidean distance is used for identifying knock. This system is implemented with a microcontroller which uses fuzzy logic controller ignition (FLIC), which aims to set proper spark advance, in accordance with operating conditions. This system can improve the engine performance for approximately 15%.

Ab initio and Molecular Dynamic models of displacement damage in crystalline and turbostratic graphite

NASA Astrophysics Data System (ADS)

McKenna, Alice

One of the functions of graphite is as a moderator in several nuclear reactor designs, including the Advanced Gas-cooled Reactor (AGR). In the reactor graphite is used to thermalise the neutrons produced in the fission reaction thus allowing a self-sustained reaction to occur. The graphite blocks, acting as the moderator, are constantly irradiated and consequently suffer damage. This thesis examines the types of damage caused using molecular dynamic (MD) simulations and ab intio calculations. Neutron damage starts with a primary knock-on atom (PKA), which is travelling so fast that it creates damage through electronic and thermal excitation (this is addressed with thermal spike simulations). When the PKA has lost energy the subsequent cascade is based on ballistic atomic displacement. These two types of simulations were performed on single crystal graphite and other carbon structures such as diamond and amorphous carbon as a comparison. The thermal spike in single crystal graphite produced results which varied from no defects to a small number of permanent defects in the structure. It is only at the high energy range that more damage is seen but these energies are less likely to occur in the nuclear reactor. The thermal spike does not create damage but it is possible that it can heal damaged sections of the graphite, which can be demonstrated with the motion of the defects when a thermal spike is applied. The cascade simulations create more damage than the thermal spike even though less energy is applied to the system. A new damage function is found with a threshold region that varies with the square root of energy in excess of the energy threshold. This is further broken down in to contributions from primary and subsequent knock-on atoms. The threshold displacement energy (TDE) is found to be Ed=25eV at 300K. In both these types of simulation graphite acts very differently to the other carbon structures. There are two types of polycrystalline graphite structures which simulations have been performed on. The difference between the two is at the grain boundaries with one having dangling bonds and the other one being bonded. The cascade showed the grain boundaries acting as a trap for the knock-on atoms which produces more damage compared with the single crystal. Finally the effects of turbostratic disorder on damage is considered. Density functional theory (DFT) was used to look at interstitials in (002) twist boundaries and how they act compared to AB stacked graphite. The results of these calculations show that the spiro interstitial is more stable in these grain boundaries, so at temperatures where the interstitial can migrate along the c direction they will segregate to (002) twist boundaries.

Relation Between Spark-Ignition Engine Knock, Detonation Waves, and Autoignition as Shown by High-Speed Photography

NASA Technical Reports Server (NTRS)

Miller, Cearcy D

1946-01-01

A critical review of literature bearing on the autoignition and detonation-wave theories of spark-ignition engine knock and on the nature of gas vibrations associated with combustion and knock results in the conclusion that neither the autoignition theory nor the detonation-wave theory is an adequate explanation of spark-ignition engine knock. A knock theory is proposed, combining the autoignition and detonation-wave theories, which introduces the idea that the detonation wave develops in autoignited or after-burning gases, and ascribes comparatively low-pitched heavy knocks to autoignition but high-pitched pinging knocks to detonation waves with the possibility of combinations of the two types of knocks. Analysis of five shots of knocking combustion, taken with the NACA high-speed motion-picture camera at the rate of 40,000 photographs per second reveals propagation speeds ranging from 3250 to more than 5500 feet per second. The range of propagation speeds from 3250 to more than 5500 feet per second is held to be considered with the proposed combined theory but not with either the simple autoignition theory or the simple detonation-wave theory.

29 CFR 1614.101 - General policy.

Code of Federal Regulations, 2010 CFR

2010-07-01

... seq.), the Age Discrimination in Employment Act (ADEA) (29 U.S.C. 621 et seq.), the Equal Pay Act (29... Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION FEDERAL SECTOR EQUAL EMPLOYMENT OPPORTUNITY Agency Program To Promote Equal Employment Opportunity § 1614.101 General policy. (a) It is the...

Cdk5 modulates cocaine reward, motivation, and striatal neuron excitability.

PubMed

Benavides, David R; Quinn, Jennifer J; Zhong, Ping; Hawasli, Ammar H; DiLeone, Ralph J; Kansy, Janice W; Olausson, Peter; Yan, Zhen; Taylor, Jane R; Bibb, James A

2007-11-21

Cyclin-dependent kinase 5 (Cdk5) regulates dopamine neurotransmission and has been suggested to serve as a homeostatic target of chronic psychostimulant exposure. To study the role of Cdk5 in the modulation of the cellular and behavioral effects of psychoactive drugs of abuse, we developed Cre/loxP conditional knock-out systems that allow temporal and spatial control of Cdk5 expression in the adult brain. Here, we report the generation of Cdk5 conditional knock-out (cKO) mice using the alphaCaMKII promoter-driven Cre transgenic line (CaMKII-Cre). In this model system, loss of Cdk5 in the adult forebrain increased the psychomotor-activating effects of cocaine. Additionally, these CaMKII-Cre Cdk5 cKO mice show enhanced incentive motivation for food as assessed by instrumental responding on a progressive ratio schedule of reinforcement. Behavioral changes were accompanied by increased excitability of medium spiny neurons in the nucleus accumbens (NAc) in Cdk5 cKO mice. To study NAc-specific effects of Cdk5, another model system was used in which recombinant adeno-associated viruses expressing Cre recombinase caused restricted loss of Cdk5 in NAc neurons. Targeted knock-out of Cdk5 in the NAc facilitated cocaine-induced locomotor sensitization and conditioned place preference for cocaine. These results suggest that Cdk5 acts as a negative regulator of neuronal excitability in the NAc and that Cdk5 may govern the behavioral effects of cocaine and motivation for reinforcement.

Development and evaluation of an efficient heterologous gene knock-in reporter system in Lactococcus lactis.

PubMed

Lu, Yifei; Yan, Hongxiang; Deng, Jiezhong; Huang, Zhigang; Jin, Xurui; Yu, Yanlan; Hu, Qiwen; Hu, Fuquan; Wang, Jing

2017-09-18

Lactococcus lactis is a food grade probiotics and widely used to express heterologous proteins. Generally, target genes are knocked into the L. lactis genome through double-crossover recombination to express heterologous proteins stably. However, creating marker-less heterologous genes knocked-in clones is laborious. In this study, an efficient heterologous gene knock-in reporter system was developed in L. lactis NZ9000. Our knock-in reporter system consists of a temperature-sensitive plasmid pJW and a recombinant L. lactis strain named NZB. The pJW contains homologous arms, and was constructed to knock-in heterologous genes at a fixed locus of NZ9000 genome. lacZ (β-galactosidase) gene was knocked into the chromosome of NZ9000 as a counter-selective marker through the plasmid pJW to generate NZB. The engineered NZB strain formed blue colonies on X-Gal plate. The desired double-crossover mutants formed white colonies distinctive from the predominantly blue colonies (parental and plasmid-integrated clones) when the embedded lacZ was replaced with the target heterologous genes carried by pJW in NZB. By using the system, the heterologous gene knocked-in clones are screened by colony phenotype change rather than by checking colonies individually. Our new knock-in reporter system provides an efficient method to create heterologous genes knocked-in clones.

29 CFR 1621.2 - Definitions.

Code of Federal Regulations, 2010 CFR

2010-07-01

... Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-THE EQUAL PAY ACT § 1621.2 Definitions. For purposes of this part, the term the Act shall mean the Equal Pay Act the Commission shall mean the Equal Employment Opportunity Commission or any of its designated representatives. ...

Gene cassette knock-in in mammalian cells and zygotes by enhanced MMEJ.

PubMed

Aida, Tomomi; Nakade, Shota; Sakuma, Tetsushi; Izu, Yayoi; Oishi, Ayu; Mochida, Keiji; Ishikubo, Harumi; Usami, Takako; Aizawa, Hidenori; Yamamoto, Takashi; Tanaka, Kohichi

2016-11-28

Although CRISPR/Cas enables one-step gene cassette knock-in, assembling targeting vectors containing long homology arms is a laborious process for high-throughput knock-in. We recently developed the CRISPR/Cas-based precise integration into the target chromosome (PITCh) system for a gene cassette knock-in without long homology arms mediated by microhomology-mediated end-joining. Here, we identified exonuclease 1 (Exo1) as an enhancer for PITCh in human cells. By combining the Exo1 and PITCh-directed donor vectors, we achieved convenient one-step knock-in of gene cassettes and floxed allele both in human cells and mouse zygotes. Our results provide a technical platform for high-throughput knock-in.

A method of estimating the knock rating of hydrocarbon fuel blend

NASA Technical Reports Server (NTRS)

Sanders, Newell D

1943-01-01

The usefulness of the knock ratings of pure hydrocarbon compounds would be increased if some reliable method of calculating the knock ratings of fuel blends was known. The purpose of this study was to investigate the possibility of developing a method of predicting the knock ratings of fuel blends.

A Method of Estimating the Knock Rating of Hydrocarbon Fuel Blends

NASA Technical Reports Server (NTRS)

Sanders, Newell D.

1943-01-01

The usefulness of the knock ratings of pure hydrocarbon compounds would be increased if some reliable method of calculating the knock ratings of fuel blends was known. The purpose of this study was to investigate the possibility of developing a method of predicting the knock ratings of fuel blends.

Porcine Knock-in Fibroblasts Expressing hDAF on α-1,3-Galactosyltransferase (GGTA1) Gene Locus.

PubMed

Kim, Ji Woo; Kim, Hye-Min; Lee, Sang Mi; Kang, Man-Jong

2012-10-01

The Galactose-α1,3-galactose (α1,3Gal) epitope is responsible for hyperacute rejection in pig-to-human xenotransplantation. Human decay-accelerating factor (hDAF) is a cell surface regulatory protein that serves as a complement inhibitor to protect self cells from complement attack. The generation of α1,3-galactosyltransferase (GGTA1) knock-out pigs expressing DAF is a necessary step for their use as organ donors for humans. In this study, we established GGTA1 knock-out cell lines expressing DAF from pig ear fibroblasts for somatic cell nuclear transfer. hDAF expression was detected in hDAF knock-in heterozygous cells, but not in normal pig cells. Expression of the GGTA1 gene was lower in the knock-in heterozygous cell line compared to the normal pig cell. Knock-in heterozygous cells afforded more effective protection against cytotoxicity with human serum than with GGTA1 knock-out heterozygous and control cells. These cell lines may be used in the production of GGTA1 knock-out and DAF expression pigs for xenotransplantation.

Simulation research on the effect of cooled EGR, supercharging and compression ratio on downsized SI engine knock

NASA Astrophysics Data System (ADS)

Shu, Gequn; Pan, Jiaying; Wei, Haiqiao; Shi, Ning

2013-03-01

Knock in spark-ignition(SI) engines severely limits engine performance and thermal efficiency. The researches on knock of downsized SI engine have mainly focused on structural design, performance optimization and advanced combustion modes, however there is little for simulation study on the effect of cooled exhaust gas recirculation(EGR) combined with downsizing technologies on SI engine performance. On the basis of mean pressure and oscillating pressure during combustion process, the effect of different levels of cooled EGR ratio, supercharging and compression ratio on engine dynamic and knock characteristic is researched with three-dimensional KIVA-3V program coupled with pressure wave equation. The cylinder pressure, combustion temperature, ignition delay timing, combustion duration, maximum mean pressure, and maximum oscillating pressure at different initial conditions are discussed and analyzed to investigate potential approaches to inhibiting engine knock while improving power output. The calculation results of the effect of just cooled EGR on knock characteristic show that appropriate levels of cooled EGR ratio can effectively suppress cylinder high-frequency pressure oscillations without obvious decrease in mean pressure. Analysis of the synergistic effect of cooled EGR, supercharging and compression ratio on knock characteristic indicates that under the condition of high supercharging and compression ratio, several times more cooled EGR ratio than that under the original condition is necessarily utilized to suppress knock occurrence effectively. The proposed method of synergistic effect of cooled EGR and downsizing technologies on knock characteristic, analyzed from the aspects of mean pressure and oscillating pressure, is an effective way to study downsized SI engine knock and provides knock inhibition approaches in practical engineering.

Influence of Compression Ratio on High Load Performance and Knock Behavior for Gasoline Port-Fuel Injection, Natural Gas Direct Injection and Blended Operation in a Spark Ignition Engine

DOE Office of Scientific and Technical Information (OSTI.GOV)

Pamminger, Michael; Sevik, James; Scarcelli, Riccardo

Natural Gas (NG) is an alternative fuel which has attracted a lot of attention recently, in particular in the US due to shale gas availability. The higher hydrogen-to-carbon (H/C) ratio, compared to gasoline, allows for decreasing carbon dioxide emissions throughout the entire engine map. Furthermore, the high knock resistance of NG allows increasing the efficiency at high engine loads compared to fuels with lower knock resistance. NG direct injection (DI) allows for fuel to be added after intake valve closing (IVC) resulting in an increase in power density compared to an injection before IVC. Steady-state engine tests were performed onmore » a single-cylinder research engine equipped with gasoline (E10) port-fuel injection (PFI) and NG DI to allow for in-cylinder blending of both fuels. Knock investigations were performed at two discrete compression ratios (CR), 10.5 and 12.5. Operating conditions span mid-load, wide-open-throttle and boosted conditions, depending on the knock response of the fuel blend. Blended operation was performed using E10 gasoline and NG. An additional gasoline type fuel (E85) with higher knock resistance than E10 was used as a high-octane reference fuel, since the octane rating of E10-NG fuel blends is unknown. Spark timing was varied at different loads under stoichiometric conditions in order to study the knock response as well as the effects on performance and efficiency. As anticipated, results suggest that the knock resistance can be increased significantly by increasing the NG amount. Comparing the engine operation with the least knock resistant fuel, E10 PFI, and the fuel blend with the highest knock resistance, 75% NG DI, shows an increase in indicated mean effective pressure of about 9 bar at CR 12.5. The usage of reference fuels with known knock characteristics allowed an assessment of knock characteristic of intermediate E10-NG blend levels. Mathematical correlations were developed allowing characterizing the occurrence of knocking combustion by using the Livengood-Wu knock integral. For most of the fueling strategies and operating conditions, the mathematical correlations show good agreement when compared to experimental data.« less

Role of the AP-5 adaptor protein complex in late endosome-to-Golgi retrieval

PubMed Central

Hirst, Jennifer; Itzhak, Daniel N.; Antrobus, Robin; Borner, Georg H. H.

2018-01-01

The AP-5 adaptor protein complex is presumed to function in membrane traffic, but so far nothing is known about its pathway or its cargo. We have used CRISPR-Cas9 to knock out the AP-5 ζ subunit gene, AP5Z1, in HeLa cells, and then analysed the phenotype by subcellular fractionation profiling and quantitative mass spectrometry. The retromer complex had an altered steady-state distribution in the knockout cells, and several Golgi proteins, including GOLIM4 and GOLM1, were depleted from vesicle-enriched fractions. Immunolocalisation showed that loss of AP-5 led to impaired retrieval of the cation-independent mannose 6-phosphate receptor (CIMPR), GOLIM4, and GOLM1 from endosomes back to the Golgi region. Knocking down the retromer complex exacerbated this phenotype. Both the CIMPR and sortilin interacted with the AP-5–associated protein SPG15 in pull-down assays, and we propose that sortilin may act as a link between Golgi proteins and the AP-5/SPG11/SPG15 complex. Together, our findings suggest that AP-5 functions in a novel sorting step out of late endosomes, acting as a backup pathway for retromer. This provides a mechanistic explanation for why mutations in AP-5/SPG11/SPG15 cause cells to accumulate aberrant endolysosomes, and highlights the role of endosome/lysosome dysfunction in the pathology of hereditary spastic paraplegia and other neurodegenerative disorders. PMID:29381698

45 CFR 287.35 - What grant amounts are available under the Personal Responsibility and Work Opportunity...

Code of Federal Regulations, 2010 CFR

2010-10-01

... NATIVE EMPLOYMENT WORKS (NEW) PROGRAM NEW Program Funding § 287.35 What grant amounts are available under the Personal Responsibility and Work Opportunity Reconciliation Act of 1996 (PRWORA) for the NEW... Personal Responsibility and Work Opportunity Reconciliation Act of 1996 (PRWORA) for the NEW Program? 287...

Expanding the horizons of disability law in India: a study from a human rights perspective.

PubMed

Chopra, Tushti

2013-01-01

Disabled/"differently abled" persons by virtue of being human have the right to enjoy human rights to life, liberty, equality, security, and dignity. However, due to social indifference, psychological barriers, a limited definition of "disability" entitling protection of law, and a lack of proper data, disabled persons in India remain an invisible category. Although several laws exit to ensure their full and effective participation in society, they remain insufficient as they are primarily based on the government's discretion. At the same time, whenever the judiciary finds an opportunity, it acts as a real protector of disabled persons, but it is not feasible to knock on the door of the judiciary for every request. Interestingly, various civil societies and human rights activists have occasionally asserted the rights of the disabled. However, unless the foundation stones of law are fortified, disabled persons cannot fully realize their rights. It is high time to enact effective laws, with timely implementation, to protect their interests and empower their capabilities that are based on a "rights-based approach" rather than on the charity, medical, or social approaches. Thus, the horizons of law must be expanded to provide a "human friendly environment" for all of the disabled to overcome the barriers that impair their development. © 2013 American Society of Law, Medicine & Ethics, Inc.

75 FR 9423 - Agency Information Collection Activities: African Growth and Opportunity Act Certificate of Origin

Federal Register 2010, 2011, 2012, 2013, 2014

2010-03-02

... Activities: African Growth and Opportunity Act Certificate of Origin AGENCY: U.S. Customs and Border... Act Certificate of Origin (AGOA). This request for comment is being made pursuant to the Paperwork... that are submitted will be summarized and included in the CBP request for Office of Management and...

76 FR 28438 - Disease, Disability, and Injury Prevention and Control Special Emphasis Panel (SEP): Initial Review

Federal Register 2010, 2011, 2012, 2013, 2014

2011-05-17

... announced below concerns ``Affordable Care Act (ACA): Childhood Obesity Research Funding Opportunity..., discussion, and evaluation of ``Affordable Care Act (ACA): Childhood Obesity Research Funding Opportunity...

76 FR 73983 - Consolidated Delegation of Authority for the Office of Fair Housing and Equal Opportunity

Federal Register 2010, 2011, 2012, 2013, 2014

2011-11-29

... Consolidated Delegation of Authority for the Office of Fair Housing and Equal Opportunity; Notices #0;#0... the Office of Fair Housing and Equal Opportunity AGENCY: Office of the Secretary, HUD. ACTION: Notice... 504 of the Rehabilitation Act of 1973, as amended; the Age Discrimination Act of 1975, as amended; and...

9 CFR 335.40 - Opportunity for presentation of views before report of criminal violations.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 9 Animals and Animal Products 2 2010-01-01 2010-01-01 false Opportunity for presentation of views... UNDER THE FEDERAL MEAT INSPECTION ACT Criminal Violations § 335.40 Opportunity for presentation of views... section, before any violation of the Federal Meat Inspection Act is reported to the Department of Justice...

Optimized knock-in of point mutations in zebrafish using CRISPR/Cas9.

PubMed

Prykhozhij, Sergey V; Fuller, Charlotte; Steele, Shelby L; Veinotte, Chansey J; Razaghi, Babak; Robitaille, Johane M; McMaster, Christopher R; Shlien, Adam; Malkin, David; Berman, Jason N

2018-06-14

We have optimized point mutation knock-ins into zebrafish genomic sites using clustered regularly interspaced palindromic repeats (CRISPR)/Cas9 reagents and single-stranded oligodeoxynucleotides. The efficiency of knock-ins was assessed by a novel application of allele-specific polymerase chain reaction and confirmed by high-throughput sequencing. Anti-sense asymmetric oligo design was found to be the most successful optimization strategy. However, cut site proximity to the mutation and phosphorothioate oligo modifications also greatly improved knock-in efficiency. A previously unrecognized risk of off-target trans knock-ins was identified that we obviated through the development of a workflow for correct knock-in detection. Together these strategies greatly facilitate the study of human genetic diseases in zebrafish, with additional applicability to enhance CRISPR-based approaches in other animal model systems.

A Photographic Study of Combustion and Knock in a Spark-Ignition Engine

NASA Technical Reports Server (NTRS)

Rothrock, A M; Spencer, R C

1938-01-01

Report presents the results of a photographic study of the combustion in a spark-ignition engine using both Schlieren and flame photographs taken at high rates of speed. Although shock waves are present after knock occurs, there was no evidence of any type of sonic or supersonic compression waves existing in the combustion gases prior to the occurrence of knock. Artificially induced shock waves in the engine did not in themselves cause knock. The photographs also indicate that, although auto-ignition ahead of the flame front may occur in conjunction with knock, it is not necessary for the occurrence of knock. There is also evidence that the reaction is not completed in the flame front but continues for some time after the flame front has passed through the charge.

76 FR 28790 - Disease, Disability, and Injury Prevention and Control Special Emphasis Panel (SEP): Initial review.

Federal Register 2010, 2011, 2012, 2013, 2014

2011-05-18

... announced below concerns Affordable Care Act (ACA): Childhood Obesity Research Funding Opportunity... ``Affordable Care Act (ACA): Childhood Obesity Research Funding Opportunity Announcement (FOA) DP11-007, Panel...

Consumer Rights and the Equal Credit Opportunity Act

ERIC Educational Resources Information Center

Griggs, Mildred B.

1978-01-01

Provisions of the Equal Credit Opportunity Act are outlined, with examples of the rights of persons seeking credit. A list of federal offices to write for information or to send complaints to is included. (MF)

The American Recovery and Reinvestment Act: Opportunities for High School Improvement

ERIC Educational Resources Information Center

Rutenberg, David

2009-01-01

The American Recovery and Reinvestment Act (ARRA) creates a unique opportunity for states, districts, and high schools to work in concert on behalf of high school students. Generally, American Recovery and Reinvestment Act funds are not intended to create new programs; rather, the majority of the funds are intended to fill gaps in existing…

Equality of Opportunity: The Making of the Americans with Disabilities Act

ERIC Educational Resources Information Center

Young, Jonathan M.

2010-01-01

In this paper the author offers a reprint of "Equality of Opportunity: The Making of the Americans with Disabilities Act" (July 26, 1997). This personal story is part and parcel of the ADA's (Americans with Disabilities Act) significance in the society. The ADA is a nondiscrimination law. It is a clarion call for transforming attitudes…

Establishment of expanded and streamlined pipeline of PITCh knock-in – a web-based design tool for MMEJ-mediated gene knock-in, PITCh designer, and the variations of PITCh, PITCh-TG and PITCh-KIKO

PubMed Central

Nakamae, Kazuki; Nishimura, Yuki; Takenaga, Mitsumasa; Sakamoto, Naoaki; Ide, Hiroshi; Sakuma, Tetsushi; Yamamoto, Takashi

2017-01-01

ABSTRACT The emerging genome editing technology has enabled the creation of gene knock-in cells easily, efficiently, and rapidly, which has dramatically accelerated research in the field of mammalian functional genomics, including in humans. We recently developed a microhomology-mediated end-joining-based gene knock-in method, termed the PITCh system, and presented various examples of its application. Since the PITCh system only requires very short microhomologies (up to 40 bp) and single-guide RNA target sites on the donor vector, the targeting construct can be rapidly prepared compared with the conventional targeting vector for homologous recombination-based knock-in. Here, we established a streamlined pipeline to design and perform PITCh knock-in to further expand the availability of this method by creating web-based design software, PITCh designer (http://www.mls.sci.hiroshima-u.ac.jp/smg/PITChdesigner/index.html), as well as presenting an experimental example of versatile gene cassette knock-in. PITCh designer can automatically design not only the appropriate microhomologies but also the primers to construct locus-specific donor vectors for PITCh knock-in. By using our newly established pipeline, a reporter cell line for monitoring endogenous gene expression, and transgenesis (TG) or knock-in/knockout (KIKO) cell line can be produced systematically. Using these new variations of PITCh, an exogenous promoter-driven gene cassette expressing fluorescent protein gene and drug resistance gene can be integrated into a safe harbor or a specific gene locus to create transgenic reporter cells (PITCh-TG) or knockout cells with reporter knock-in (PITCh-KIKO), respectively. PMID:28453368

Establishment of expanded and streamlined pipeline of PITCh knock-in - a web-based design tool for MMEJ-mediated gene knock-in, PITCh designer, and the variations of PITCh, PITCh-TG and PITCh-KIKO.

PubMed

Nakamae, Kazuki; Nishimura, Yuki; Takenaga, Mitsumasa; Nakade, Shota; Sakamoto, Naoaki; Ide, Hiroshi; Sakuma, Tetsushi; Yamamoto, Takashi

2017-05-04

The emerging genome editing technology has enabled the creation of gene knock-in cells easily, efficiently, and rapidly, which has dramatically accelerated research in the field of mammalian functional genomics, including in humans. We recently developed a microhomology-mediated end-joining-based gene knock-in method, termed the PITCh system, and presented various examples of its application. Since the PITCh system only requires very short microhomologies (up to 40 bp) and single-guide RNA target sites on the donor vector, the targeting construct can be rapidly prepared compared with the conventional targeting vector for homologous recombination-based knock-in. Here, we established a streamlined pipeline to design and perform PITCh knock-in to further expand the availability of this method by creating web-based design software, PITCh designer ( http://www.mls.sci.hiroshima-u.ac.jp/smg/PITChdesigner/index.html ), as well as presenting an experimental example of versatile gene cassette knock-in. PITCh designer can automatically design not only the appropriate microhomologies but also the primers to construct locus-specific donor vectors for PITCh knock-in. By using our newly established pipeline, a reporter cell line for monitoring endogenous gene expression, and transgenesis (TG) or knock-in/knockout (KIKO) cell line can be produced systematically. Using these new variations of PITCh, an exogenous promoter-driven gene cassette expressing fluorescent protein gene and drug resistance gene can be integrated into a safe harbor or a specific gene locus to create transgenic reporter cells (PITCh-TG) or knockout cells with reporter knock-in (PITCh-KIKO), respectively.

Bowlegs and Knock-Knees

MedlinePlus

... Español Text Size Email Print Share Bowlegs and Knock-Knees Page Content Article Body Toddlers’ legs often ... about two years old, then they’ll look knock-kneed until they are about six years of ...

76 FR 50284 - Request for Public Comments on Interim Review of Eligibility of Cote d'Ivoire, Guinea, and Niger...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-08-12

... Eligibility of Cote d'Ivoire, Guinea, and Niger for Benefits Under the African Growth and Opportunity Act.... SUMMARY: The African Growth and Opportunity Act Implementation Subcommittee of the Trade Policy Staff... eligibility of Cote d'Ivoire, Guinea, and Niger to receive the benefits of the African Growth and Opportunity...

Leveraging Community Colleges in the Workforce Innovation and Opportunity Act: A Blueprint for State Policymakers. State-Federal Partnerships in Postsecondary Education

ERIC Educational Resources Information Center

Campbell, Colleen; Love, Ivy

2016-01-01

The Workforce Innovation and Opportunity Act (WIOA) of 2014 offers multiple opportunities to align the workforce development efforts of these stakeholders through structural measures and targeted support. In this paper, the authors examine ways that WIOA can influence a state's job training environment and highlight the crucial role of community…

Objective evaluation of the knocking sound of a diesel engine considering the temporal and frequency masking effect simultaneously

NASA Astrophysics Data System (ADS)

Yun, Dong-Un; Lee, Sang-Kwon

2017-06-01

In this paper, we present a novel method for an objective evaluation of knocking noise emitted by diesel engines based on the temporal and frequency masking theory. The knocking sound of a diesel engine is a vibro-acoustic sound correlated with the high-frequency resonances of the engine structure and a periodic impulsive sound with amplitude modulation. Its period is related to the engine speed and includes specific frequency bands related to the resonances of the engine structure. A knocking sound with the characteristics of a high-frequency impulsive wave can be masked by low-frequency sounds correlated with the harmonics of the firing frequency and broadband noise. The degree of modulation of the knocking sound signal was used for such objective evaluations in previous studies, without considering the masking effect. However, the frequency masking effect must be considered for the objective evaluation of the knocking sound. In addition to the frequency masking effect, the temporal masking effect occurs because the period of the knocking sound changes according to the engine speed. Therefore, an evaluation method considering the temporal and frequency masking effect is required to analyze the knocking sound objectively. In this study, an objective evaluation method considering the masking effect was developed based on the masking theory of sound and signal processing techniques. The method was applied successfully for the objective evaluation of the knocking sound of a diesel engine.

Public Notice: Notice of Proposed Administrative Penalty Assessments and Opportunity to Comment

EPA Pesticide Factsheets

EPA is providing notice of a proposed administrative penalty assessment for alleged violations of the Clean Water Act (Act). EPA is also providing notice of opportunity to comment on the proposed penalty assessment.

Ketone Body Metabolic Enzyme OXCT1 Regulates Prostate Cancer Chemoresistance

DTIC Science & Technology

2015-12-01

increased ADP/ATP, NAD +/NADH and oxygen consumption in docetaxel treated cells compared to control knock down cells, therefore induced metabolic...substrate for mitochondrial oxidative phosphorylation and ATP biosynthesis. Next, we examined NAD +/NADH levels in OXC1 knock down prostate cancer cells...The results showed that after docetaxel treatment, NAD + level was significantly increased in OXCT1 knock down cells compared to control knock down

Knock probability estimation through an in-cylinder temperature model with exogenous noise

NASA Astrophysics Data System (ADS)

Bares, P.; Selmanaj, D.; Guardiola, C.; Onder, C.

2018-01-01

This paper presents a new knock model which combines a deterministic knock model based on the in-cylinder temperature and an exogenous noise disturbing this temperature. The autoignition of the end-gas is modelled by an Arrhenius-like function and the knock probability is estimated by propagating a virtual error probability distribution. Results show that the random nature of knock can be explained by uncertainties at the in-cylinder temperature estimation. The model only has one parameter for calibration and thus can be easily adapted online. In order to reduce the measurement uncertainties associated with the air mass flow sensor, the trapped mass is derived from the in-cylinder pressure resonance, which improves the knock probability estimation and reduces the number of sensors needed for the model. A four stroke SI engine was used for model validation. By varying the intake temperature, the engine speed, the injected fuel mass, and the spark advance, specific tests were conducted, which furnished data with various knock intensities and probabilities. The new model is able to predict the knock probability within a sufficient range at various operating conditions. The trapped mass obtained by the acoustical model was compared in steady conditions by using a fuel balance and a lambda sensor and differences below 1 % were found.

Development of versatile non-homologous end joining-based knock-in module for genome editing.

PubMed

Sawatsubashi, Shun; Joko, Yudai; Fukumoto, Seiji; Matsumoto, Toshio; Sugano, Shigeo S

2018-01-12

CRISPR/Cas9-based genome editing has dramatically accelerated genome engineering. An important aspect of genome engineering is efficient knock-in technology. For improved knock-in efficiency, the non-homologous end joining (NHEJ) repair pathway has been used over the homology-dependent repair pathway, but there remains a need to reduce the complexity of the preparation of donor vectors. We developed the versatile NHEJ-based knock-in module for genome editing (VIKING). Using the consensus sequence of the time-honored pUC vector to cut donor vectors, any vector with a pUC backbone could be used as the donor vector without customization. Conditions required to minimize random integration rates of the donor vector were also investigated. We attempted to isolate null lines of the VDR gene in human HaCaT keratinocytes using knock-in/knock-out with a selection marker cassette, and found 75% of clones isolated were successfully knocked-in. Although HaCaT cells have hypotetraploid genome composition, the results suggest multiple clones have VDR null phenotypes. VIKING modules enabled highly efficient knock-in of any vectors harboring pUC vectors. Users now can insert various existing vectors into an arbitrary locus in the genome. VIKING will contribute to low-cost genome engineering.

Tracking of Pacific walruses in the Chukchi Sea using a single hydrophone.

PubMed

Mouy, Xavier; Hannay, David; Zykov, Mikhail; Martin, Bruce

2012-02-01

The vocal repertoire of Pacific walruses includes underwater sound pulses referred to as knocks and bell-like calls. An extended acoustic monitoring program was performed in summer 2007 over a large region of the eastern Chukchi Sea using autonomous seabed-mounted acoustic recorders. Walrus knocks were identified in many of the recordings and most of these sounds included multiple bottom and surface reflected signals. This paper investigates the use of a localization technique based on relative multipath arrival times (RMATs) for potential behavior studies. First, knocks are detected using a semi-automated kurtosis-based algorithm. Then RMATs are matched to values predicted by a ray-tracing model. Walrus tracks with vertical and horizontal movements were obtained. The tracks included repeated dives between 4.0 m and 15.5 m depth and a deep dive to the sea bottom (53 m). Depths at which bell-like sounds are produced, average knock production rate and source levels estimates of the knocks were determined. Bell sounds were produced at all depths throughout the dives. Average knock production rates varied from 59 to 75 knocks/min. Average source level of the knocks was estimated to 177.6 ± 7.5 dB re 1 μPa peak @ 1 m. © 2012 Acoustical Society of America

Engineered Coatings Inc.; Proposed Settlement of Clean Water Act Class I Administrative Penalty and Opportunity to Comment

EPA Pesticide Factsheets

Public Notice of Proposed Settlement of Clean Water Act Class I Administrative Penalty and Opportunity to Comment In the Matter of Engineered Coatings Inc., 3154 Harcourt Street, Compton, California, 90221.

29 CFR 1621.1 - Purpose.

Code of Federal Regulations, 2010 CFR

2010-07-01

... to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-THE EQUAL PAY ACT § 1621.1 Purpose. The regulations set forth in this part contain the procedures established by the Equal Employment Opportunity Commission for issuing opinion letters under the Equal Pay Act. ...

77 FR 25168 - Sunshine Act Meeting

Federal Register 2010, 2011, 2012, 2013, 2014

2012-04-27

... EQUAL EMPLOYMENT OPPORTUNITY COMMISSION Sunshine Act Meeting AGENCY HOLDING THE MEETING: Equal Employment Opportunity Commission. FEDERAL REGISTER CITATION OF PREVIOUS ANNOUNCEMENT: 77 FR 24201, Monday.... Enforcement Guidance on the Consideration of Arrest and Conviction Records in Employment Decisions under Title...

RS-1 enhances CRISPR/Cas9- and TALEN-mediated knock-in efficiency.

PubMed

Song, Jun; Yang, Dongshan; Xu, Jie; Zhu, Tianqing; Chen, Y Eugene; Zhang, Jifeng

2016-01-28

Zinc-finger nuclease, transcription activator-like effector nuclease and CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated protein 9) are becoming major tools for genome editing. Importantly, knock-in in several non-rodent species has been finally achieved thanks to these customizable nucleases; yet the rates remain to be further improved. We hypothesize that inhibiting non-homologous end joining (NHEJ) or enhancing homology-directed repair (HDR) will improve the nuclease-mediated knock-in efficiency. Here we show that the in vitro application of an HDR enhancer, RS-1, increases the knock-in efficiency by two- to five-fold at different loci, whereas NHEJ inhibitor SCR7 has minimal effects. We then apply RS-1 for animal production and have achieved multifold improvement on the knock-in rates as well. Our work presents tools to nuclease-mediated knock-in animal production, and sheds light on improving gene-targeting efficiencies on pluripotent stem cells.

'Knock, and it shall be opened': knocking out and knocking in to reveal mechanisms of disease and novel therapies.

PubMed

Hacking, Douglas F

2008-12-01

Recent significant advances in molecular biology have generated genetically modified bacteria, yeast, nematodes, fruit flies, and fish. However, it is the genetic modification of mammalian model organisms, particularly the mouse, that has the greatest potential to shed light on human development, physiology and pathology in ways that have significant implications for neonatal and paediatric clinical practice. Here, we review some of the techniques for knocking out (inactivating), mutating and knocking in (inserting) selected genes that are important to neonatology and show how this research will lead both to a better understanding of disease and to novel therapies for infants and children.

Hige Compression Ratio Turbo Gasoline Engine Operation Using Alcohol Enhancement

DOE Office of Scientific and Technical Information (OSTI.GOV)

Heywood, John; Jo, Young Suk; Lewis, Raymond

The overall objective of this project was to quantify the potential for improving the performance and efficiency of gasoline engine technology by use of alcohols to suppress knock. Knock-free operation is obtained by direct injection of a second “anti-knock” fuel such as ethanol, which suppresses knock when, with gasoline fuel, knock would occur. Suppressing knock enables increased turbocharging, engine downsizing, and use of higher compression ratios throughout the engine’s operating map. This project combined engine testing and simulation to define knock onset conditions, with different mixtures of gasoline and alcohol, and with this information quantify the potential for improving themore » efficiency of turbocharged gasoline spark-ignition engines, and the on-vehicle fuel consumption reductions that could then be realized. The more focused objectives of this project were therefore to: Determine engine efficiency with aggressive turbocharging and downsizing and high compression ratio (up to a compression ratio of 13.5:1) over the engine’s operating range; Determine the knock limits of a turbocharged and downsized engine as a function of engine speed and load; Determine the amount of the knock-suppressing alcohol fuel consumed, through the use of various alcohol-gasoline and alcohol-water gasoline blends, for different driving cycles, relative to the gasoline consumed; Determine implications of using alcohol-boosted engines, with their higher efficiency operation, in both light-duty and medium-duty vehicle sectors.« less

The Income and Jobs Action Act of 1985. Hearing before the Subcommittee on Employment Opportunities of the Committee on Education and Labor, House of Representatives, Ninety-Ninth Congress, First Session, on H.R. 1398. (Chicago, IL, September 4, 1985).

ERIC Educational Resources Information Center

Congress of the U.S., Washington, DC. House Committee on Education and Labor.

The proceedings of a House Subcommittee on Employment Opportunities hearing, held in Chicago, are provided in this document. Testimony is presented on a proposed bill, H.R. 1938 (the Income and Jobs Action Act), which aims at reducing unemployment by fully implementing the Employment Act of 1946 and the Full Employment and Balanced Growth Act of…

29 CFR 1625.1 - Definitions.

Code of Federal Regulations, 2012 CFR

2012-07-01

... Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION AGE DISCRIMINATION IN EMPLOYMENT ACT Interpretations § 1625.1 Definitions. The Equal Employment Opportunity Commission is hereinafter referred to as... have the meanings set forth in section 11 of the Age Discrimination in Employment Act of 1967, as...

29 CFR 1625.1 - Definitions.

Code of Federal Regulations, 2014 CFR

2014-07-01

... Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION AGE DISCRIMINATION IN EMPLOYMENT ACT Interpretations § 1625.1 Definitions. The Equal Employment Opportunity Commission is hereinafter referred to as... have the meanings set forth in section 11 of the Age Discrimination in Employment Act of 1967, as...

29 CFR 1625.1 - Definitions.

Code of Federal Regulations, 2010 CFR

2010-07-01

... Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION AGE DISCRIMINATION IN EMPLOYMENT ACT Interpretations § 1625.1 Definitions. The Equal Employment Opportunity Commission is hereinafter referred to as... have the meanings set forth in section 11 of the Age Discrimination in Employment Act of 1967, as...

29 CFR 1625.1 - Definitions.

Code of Federal Regulations, 2011 CFR

2011-07-01

... Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION AGE DISCRIMINATION IN EMPLOYMENT ACT Interpretations § 1625.1 Definitions. The Equal Employment Opportunity Commission is hereinafter referred to as... have the meanings set forth in section 11 of the Age Discrimination in Employment Act of 1967, as...

29 CFR 1625.1 - Definitions.

Code of Federal Regulations, 2013 CFR

2013-07-01

... Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION AGE DISCRIMINATION IN EMPLOYMENT ACT Interpretations § 1625.1 Definitions. The Equal Employment Opportunity Commission is hereinafter referred to as... have the meanings set forth in section 11 of the Age Discrimination in Employment Act of 1967, as...

78 FR 16908 - Determinations Under the African Growth and Opportunity Act

Federal Register 2010, 2011, 2012, 2013, 2014

2013-03-19

... STATES TRADE REPRESENTATIVE Determinations Under the African Growth and Opportunity Act AGENCY... substantial progress toward implementing and following, the customs procedures required by the African Growth... lesser developed beneficiary sub-Saharan African country. In Proclamation 7350 (October 2, 2000), the...

75 FR 48306 - Submission for OMB Review; Comment Request

Federal Register 2010, 2011, 2012, 2013, 2014

2010-08-10

... Opportunities to Meaningfully Promote Excellence in Technology, Education and Science Act, Pub. L. 110-69), which seeks to strengthen education and research related to science and technology (``America Creating Opportunities to Meaningfully Promote Excellence in Technology, Education, and Science Act''). This legislation...

Determination of knock characteristics in spark ignition engines: an approach based on ensemble empirical mode decomposition

NASA Astrophysics Data System (ADS)

Li, Ning; Yang, Jianguo; Zhou, Rui; Liang, Caiping

2016-04-01

Knock is one of the major constraints to improve the performance and thermal efficiency of spark ignition (SI) engines. It can also result in severe permanent engine damage under certain operating conditions. Based on the ensemble empirical mode decomposition (EEMD), this paper proposes a new approach to determine the knock characteristics in SI engines. By adding a uniformly distributed and finite white Gaussian noise, the EEMD can preserve signal continuity in different scales and therefore alleviates the mode-mixing problem occurring in the classic empirical mode decomposition (EMD). The feasibilities of applying the EEMD to detect the knock signatures of a test SI engine via the pressure signal measured from combustion chamber and the vibration signal measured from cylinder head are investigated. Experimental results show that the EEMD-based method is able to detect the knock signatures from both the pressure signal and vibration signal, even in initial stage of knock. Finally, by comparing the application results with those obtained by short-time Fourier transform (STFT), Wigner-Ville distribution (WVD) and discrete wavelet transform (DWT), the superiority of the EEMD method in determining knock characteristics is demonstrated.

Relation between Spark-Ignition Engine Knock, Detonation Waves, and Autoignition as Shown by High-Speed Photography

DTIC Science & Technology

1946-01-01

unfortunate” fiat this work has not, in the past few years , received more carcf~ll considera- tion. Th~ photographs of %kolik and Voinov wcro taken through a...with the propo8ed combined theory but not with either the simple autoignition theory or the simple detonation- wace theory. INTRODUCTION Knock is one of...countries for about 25 yeara. The past researches on knock have uncovered an immense amount of information, not only concerning the basic nature of knock but

System and method for controlling engine knock using electro-hydraulic valve actuation

DOEpatents

Brennan, Daniel G

2013-12-10

A control system for an engine includes a knock control module and a valve control module. The knock control module adjusts a period that one or more of an intake valve and an exhaust valve of a cylinder are open based on engine knock corresponding to the cylinder. The valve control module, based on the adjusted period, controls the one or more of the intake valve and the exhaust valve using one or more hydraulic actuators.

Understanding fuel anti-knock performances in modern SI engines using fundamental HCCI experiments

DOE PAGES

Yang, Yi; Dec, John E.; Sjoberg, Magnus; ...

2015-08-19

Modern spark-ignition (SI) engine technologies have considerably changed in-cylinder conditions under which fuel autoignition and engine knock take place. In this paper, fundamental HCCI engine experiments are proposed as a means for characterizing the impact of these technologies on the knock propensity of different fuels. In particular, the impacts of turbocharging, direct injection (DI), and downspeeding on operation with ethanol and gasoline are investigated to demonstrate this approach. Results reported earlier for ethanol and gasoline on HCCI combustion are revisited with the new perspective of how their autoignition characteristics fit into the anti-knock requirement in modern SI engines. For example,more » the weak sensitivity to pressure boost demonstrated by ethanol in HCCI autoignition can be used to explain the strong knock resistance of ethanol fuels for turbocharged SI engines. Further, ethanol's high sensitivity to charge temperature makes charge cooling, which can be produced by fuel vaporization via direct injection or by piston expansion via spark-timing retard, very effective for inhibiting knock. On the other hand, gasoline autoignition shows a higher sensitivity to pressure, so only very low pressure boost can be applied before knock occurs. Gasoline also demonstrates low temperature sensitivity, so it is unable to make as effective use of the charge cooling produced by fuel vaporization or spark retard. These arguments comprehensively explain literature results on ethanol's substantially better anti-knock performance over gasoline in modern turbocharged DISI engines. Fundamental HCCI experiments such as these can thus be used as a diagnostic and predictive tool for knock-limited SI engine performance for various fuels. As a result, examples are presented where HCCI experiments are used to identify biofuel compounds with good potential for modern SI-engine applications.« less

Deficiency in the manganese efflux transporter SLC30A10 induces severe hypothyroidism in mice.

PubMed

Hutchens, Steven; Liu, Chunyi; Jursa, Thomas; Shawlot, William; Chaffee, Beth K; Yin, Weiling; Gore, Andrea C; Aschner, Michael; Smith, Donald R; Mukhopadhyay, Somshuvra

2017-06-09

Manganese is an essential metal that becomes toxic at elevated levels. Loss-of-function mutations in SLC30A10, a cell-surface-localized manganese efflux transporter, cause a heritable manganese metabolism disorder resulting in elevated manganese levels and parkinsonian-like movement deficits. The underlying disease mechanisms are unclear; therefore, treatment is challenging. To understand the consequences of loss of SLC30A10 function at the organism level, we generated Slc30a10 knock-out mice. During early development, knock-outs were indistinguishable from controls. Surprisingly, however, after weaning and compared with controls, knock-out mice failed to gain weight, were smaller, and died prematurely (by ∼6-8 weeks of age). At 6 weeks, manganese levels in the brain, blood, and liver of the knock-outs were ∼20-60-fold higher than controls. Unexpectedly, histological analyses revealed that the brain and liver of the knock-outs were largely unaffected, but their thyroid exhibited extensive alterations. Because hypothyroidism leads to growth defects and premature death in mice, we assayed for changes in thyroid and pituitary hormones. At 6 weeks and compared with controls, the knock-outs had markedly reduced thyroxine levels (∼50-80%) and profoundly increased thyroid-stimulating hormone levels (∼800-1000-fold), indicating that Slc30a10 knock-out mice develop hypothyroidism. Importantly, a low-manganese diet produced lower tissue manganese levels in the knock-outs and rescued the phenotype, suggesting that manganese toxicity was the underlying cause. Our unanticipated discovery highlights the importance of determining the role of thyroid dysfunction in the onset and progression of manganese-induced disease and identifies Slc30a10 knock-out mice as a new model for studying thyroid biology. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Knock-in mice harboring a Ca(2+) desensitizing mutation in cardiac troponin C develop early onset dilated cardiomyopathy.

PubMed

McConnell, Bradley K; Singh, Sonal; Fan, Qiying; Hernandez, Adriana; Portillo, Jesus P; Reiser, Peter J; Tikunova, Svetlana B

2015-01-01

The physiological consequences of aberrant Ca(2+) binding and exchange with cardiac myofilaments are not clearly understood. In order to examine the effect of decreasing Ca(2+) sensitivity of cTnC on cardiac function, we generated knock-in mice carrying a D73N mutation (not known to be associated with heart disease in human patients) in cTnC. The D73N mutation was engineered into the regulatory N-domain of cTnC in order to reduce Ca(2+) sensitivity of reconstituted thin filaments by increasing the rate of Ca(2+) dissociation. In addition, the D73N mutation drastically blunted the extent of Ca(2+) desensitization of reconstituted thin filaments induced by cTnI pseudo-phosphorylation. Compared to wild-type mice, heterozygous knock-in mice carrying the D73N mutation exhibited a substantially decreased Ca(2+) sensitivity of force development in skinned ventricular trabeculae. Kaplan-Meier survival analysis revealed that median survival time for knock-in mice was 12 weeks. Echocardiographic analysis revealed that knock-in mice exhibited increased left ventricular dimensions with thinner walls. Echocardiographic analysis also revealed that measures of systolic function, such as ejection fraction (EF) and fractional shortening (FS), were dramatically reduced in knock-in mice. In addition, knock-in mice displayed electrophysiological abnormalities, namely prolonged QRS and QT intervals. Furthermore, ventricular myocytes isolated from knock-in mice did not respond to β-adrenergic stimulation. Thus, knock-in mice developed pathological features similar to those observed in human patients with dilated cardiomyopathy (DCM). In conclusion, our results suggest that decreasing Ca(2+) sensitivity of the regulatory N-domain of cTnC is sufficient to trigger the development of DCM.

29 CFR 1614.503 - Enforcement of final Commission decisions.

Code of Federal Regulations, 2010 CFR

2010-07-01

... ADEA, the Equal Pay Act or the Rehabilitation Act and to seek judicial review of the agency's refusal... 1614.503 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION FEDERAL SECTOR EQUAL EMPLOYMENT OPPORTUNITY Remedies and Enforcement § 1614.503 Enforcement of final...

28 CFR 42.613 - Definitions.

Code of Federal Regulations, 2010 CFR

2010-07-01

... Pay Act and by title VI, title IX, or the revenue sharing act. (g) Recipient means any State... Judicial Administration DEPARTMENT OF JUSTICE NONDISCRIMINATION; EQUAL EMPLOYMENT OPPORTUNITY; POLICIES AND...) Chairman of the EEOC refers to the Chairman of the Equal Employment Opportunity Commission, or his or her...

78 FR 2397 - Sunshine Act Notice

Federal Register 2010, 2011, 2012, 2013, 2014

2013-01-11

... EQUAL EMPLOYMENT OPPORTUNITY COMMISSION Sunshine Act Notice AGENCY HOLDING THE MEETING: Equal Employment Opportunity Commission. DATE AND TIME: Wednesday, January 16, 2013, 9:30 a.m. Eastern Time. PLACE: Commission Meeting Room on the First Floor of the EEOC Office Building, 131 ``M'' Street NE., Washington, DC...

78 FR 16501 - Sunshine Act Meeting Notice

Federal Register 2010, 2011, 2012, 2013, 2014

2013-03-15

... EQUAL EMPLOYMENT OPPORTUNITY COMMISSION Sunshine Act Meeting Notice AGENCY: Equal Employment Opportunity Commission. DATE AND TIME: Wednesday, March 20, 2013, 9:50 a.m. Eastern Time. PLACE: Commission Meeting Room on the First Floor of the EEOC Office Building, 131 ``M'' Street NE., Washington, DC 20507...

Generation of an allelic series of knock-in mice using recombinase-mediated cassette exchange (RMCE).

PubMed

Roebroek, Anton J M; Van Gool, Bart

2014-01-01

Molecular genetic strategies applying embryonic stem cell (ES cell) technologies to study the function of a gene in mice or to generate a mouse model for a human disease are continuously under development. Next to (conditional) inactivation of genes the application and importance of approaches to generate knock-in mutations are increasing. In this chapter the principle and application of recombinase-mediated cassette exchange (RMCE) are discussed as being a new emerging knock-in strategy, which enables easy generation of a series of different knock-in mutations within one gene. An RMCE protocol, which was used to generate a series of different knock-in mutations in the Lrp1 gene of ES cells, is described in detail as an example of how RMCE can be used to generate highly efficiently an allelic series of differently modified ES cell clones from a parental modified ES cell clone. Subsequently the differently modified ES cell clones can be used to generate an allelic series of mutant knock-in mice.

RS-1 enhances CRISPR/Cas9- and TALEN-mediated knock-in efficiency

PubMed Central

Song, Jun; Yang, Dongshan; Xu, Jie; Zhu, Tianqing; Chen, Y. Eugene; Zhang, Jifeng

2016-01-01

Zinc-finger nuclease, transcription activator-like effector nuclease and CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated protein 9) are becoming major tools for genome editing. Importantly, knock-in in several non-rodent species has been finally achieved thanks to these customizable nucleases; yet the rates remain to be further improved. We hypothesize that inhibiting non-homologous end joining (NHEJ) or enhancing homology-directed repair (HDR) will improve the nuclease-mediated knock-in efficiency. Here we show that the in vitro application of an HDR enhancer, RS-1, increases the knock-in efficiency by two- to five-fold at different loci, whereas NHEJ inhibitor SCR7 has minimal effects. We then apply RS-1 for animal production and have achieved multifold improvement on the knock-in rates as well. Our work presents tools to nuclease-mediated knock-in animal production, and sheds light on improving gene-targeting efficiencies on pluripotent stem cells. PMID:26817820

Workplace Discrimination and Visual Impairment: A Comparison of Equal Employment Opportunity Commission Charges and Resolutions under the Americans with Disabilities Act and Americans with Disabilities Amendments Act

ERIC Educational Resources Information Center

Victor, Callie M.; Thacker, Leroy R.; Gary, Kelli W.; Pawluk, Dianne T. V.; Copolillo, Al

2017-01-01

To guarantee equal opportunities and treatment in employment for individuals with disabilities, Congress enacted Title I of the Americans with Disabilities Act (ADA) in 1992. This law states, "No covered entity shall discriminate against a qualified individual on the basis of disability in regard to job application procedures, the hiring,…

Role of blood ribosomal protein S19 in coagulum resorption: a study using Gln137Glu-ribosomal protein S19 gene knock-in mouse.

PubMed

Chen, Jun; Fujino, Rika; Zhao, Rui; Semba, Umeko; Araki, Kimi; Yamamoto, Tetsuro

2014-11-01

Sera of human, guinea pig or mouse contain a strong monocyte chemoattractant capacity that is attributed to the ribosomal protein S19 (RP S19) oligomers generated during blood coagulation. In contrast, sera prepared from Gln137Glu-RP S19 gene knock-in mice contained negligible chemoattractant capacity. When coagula that had been pre-formed from the blood of both the wild type and knock-in mice were intraperitoneally inserted into host mice, after 3 days of recovery, the knock-in mouse coagula remained larger than the wild type mouse coagula. The wild type mouse coagula were covered by multiple macrophage layers at the surface and were infiltrated inside by macrophages. Knock-in mouse coagula exhibited less macrophage involvement. When coagula of knock-in mice and coagula of knock-in mice containing C5a/RP S19, an artificial substitute of the RP S19 oligomers, were intraperitoneally inserted as pairs, the C5a/RP S19 containing coagulum was more rapidly absorbed, concomitant with increased macrophage involvement. Finally, when the knock-in mouse and wild type mouse coagula pairs were inserted into mice in which macrophages had been depleted using clodronate liposome, the size difference of recovered coagula was reversed. These results indicate the importance of the RP S19 oligomer-induced macrophage recruitment in coagulum resorption. © 2014 Japanese Society of Pathology and Wiley Publishing Asia Pty Ltd.

Overexpression of ZIC5 promotes proliferation in non-small cell lung cancer

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sun, Qi; Shi, Run; Wang, Xin

Background: Non-small cell lung cancer (NSCLC) has become the leading cause of cancer-related deaths. It is therefore urgent that we identify new molecular targets to help cure NSCLC patients. Here, we identified ZIC5 as a potential novel oncogene. Methods: We detected the expression of ZIC5 in tumor and normal tissues of NSCLC patients using quantitative real-time PCR and explored its clinical appearance. We then knocked down ZIC5 to observe changes in NSCLC cell proliferation and metastasis. Nude mouse xenograft models were established to measure ZIC5's function in vivo. Results: Our results revealed that ZIC5 was expressed at dramatically higher levels inmore » NSCLC tumor tissues than in normal tissues. High levels of ZIC5 expression were associated with a higher primary tumor grade. ZIC5 expression was significantly inhibited by small interfering RNA. After silencing ZIC5, the metastatic capacity of NSCLC cells was clearly lower. Knocking down ZIC5 significantly inhibited the proliferation of NSCLC cells, causing the cell cycle to be arrested in G2 phase. Xenograft tumor models showed that knocking down ZIC5 also inhibited tumor growth in vivo. Q-PCR and western blot analysis revealed that ZIC5 expression was closely associated with CCNB1 and CDK1 complex expression, while other cell cycle-related genes showed no significant correlation with ZIC5. Conclusions: Our experiment show that ZIC5 is highly upregulated in NSCLC tumor tissues and suggest that ZIC5 may act as an oncogene by influencing CCNB1 and CDK1 complex expression. ZIC5 may therefore be a potential biomarker and therapeutic target for NSCLC patients.« less

Leptin signaling in GABA neurons, but not glutamate neurons, is required for reproductive function.

PubMed

Zuure, Wieteke A; Roberts, Amy L; Quennell, Janette H; Anderson, Greg M

2013-11-06

The adipocyte-derived hormone leptin acts in the brain to modulate the central driver of fertility: the gonadotropin releasing hormone (GnRH) neuronal system. This effect is indirect, as GnRH neurons do not express leptin receptors (LEPRs). Here we test whether GABAergic or glutamatergic neurons provide the intermediate pathway between the site of leptin action and the GnRH neurons. Leptin receptors were deleted from GABA and glutamate neurons using Cre-Lox transgenics, and the downstream effects on puberty onset and reproduction were examined. Both mouse lines displayed the expected increase in body weight and region-specific loss of leptin signaling in the hypothalamus. The GABA neuron-specific LEPR knock-out females and males showed significantly delayed puberty onset. Adult fertility observations revealed that these knock-out animals have decreased fecundity. In contrast, glutamate neuron-specific LEPR knock-out mice displayed normal fertility. Assessment of the estrogenic hypothalamic-pituitary-gonadal axis regulation in females showed that leptin action on GABA neurons is not necessary for estradiol-mediated suppression of tonic luteinizing hormone secretion (an indirect measure of GnRH neuron activity) but is required for regulation of a full preovulatory-like luteinizing hormone surge. In conclusion, leptin signaling in GABAergic (but not glutamatergic neurons) plays a critical role in the timing of puberty onset and is involved in fertility regulation throughout adulthood in both sexes. These results form an important step in explaining the role of central leptin signaling in the reproductive system. Limiting the leptin-to-GnRH mediators to GABAergic cells will enable future research to focus on a few specific types of neurons.

Overlap of heritable influences between cannabis use disorder, frequency of use and opportunity to use cannabis: trivariate twin modelling and implications for genetic design.

PubMed

Hines, Lindsey A; Morley, Katherine I; Rijsdijk, Fruhling; Strang, John; Agrawal, Arpana; Nelson, Elliot C; Statham, Dixie; Martin, Nicholas G; Lynskey, Michael T

2018-03-13

The genetic component of Cannabis Use Disorder may overlap with influences acting more generally on early stages of cannabis use. This paper aims to determine the extent to which genetic influences on the development of cannabis abuse/dependence are correlated with those acting on the opportunity to use cannabis and frequency of use. A cross-sectional study of 3303 Australian twins, measuring age of onset of cannabis use opportunity, lifetime frequency of cannabis use, and lifetime DSM-IV cannabis abuse/dependence. A trivariate Cholesky decomposition estimated additive genetic (A), shared environment (C) and unique environment (E) contributions to the opportunity to use cannabis, the frequency of cannabis use, cannabis abuse/dependence, and the extent of overlap between genetic and environmental factors associated with each phenotype. Variance components estimates were A = 0.64 [95% confidence interval (CI) 0.58-0.70] and E = 0.36 (95% CI 0.29-0.42) for age of opportunity to use cannabis, A = 0.74 (95% CI 0.66-0.80) and E = 0.26 (95% CI 0.20-0.34) for cannabis use frequency, and A = 0.78 (95% CI 0.65-0.88) and E = 0.22 (95% CI 0.12-0.35) for cannabis abuse/dependence. Opportunity shares 45% of genetic influences with the frequency of use, and only 17% of additive genetic influences are unique to abuse/dependence from those acting on opportunity and frequency. There are significant genetic contributions to lifetime cannabis abuse/dependence, but a large proportion of this overlaps with influences acting on opportunity and frequency of use. Individuals without drug use opportunity are uninformative, and studies of drug use disorders must incorporate individual exposure to accurately identify aetiology.

Bioethics as several kinds of writing.

PubMed

Nelson, J L

1999-04-01

Three different models are described of the relationship of bioethics to the press. The first two are familiar: bioethicists often are interviewed by journalists seeking background and short quotes to insert in a story; alternately, bioethicists sometimes themselves act as journalists of a sort, writing op-eds, articles or even longer works designed for wide readership. These models share the notion that bioethicists can provide information and ideas that increase the quality of people's thinking on moral matters. They share also a common difficulty: do the constraints the media impose on bioethical discourse keep bioethicists from deepening public reflection, and if not, how can those constraints be most effectively kept from distorting what bioethicists wish to say? The third model reverses--in part--the presupposition that bioethics bestows moral sophistication on a public naive about ethical issues, holding rather that matters run both ways; bioethicists stand to learn a great deal from their interactions with various publics and the media that serve them. On this view, the constraints imposed by media conventions constitute opportunities for new and potentially important forms of bioethical writing. Various concerns generated by the first two models are surveyed. It is concluded that while none of the difficulties constitute knock-down arguments against these forms of collaborating with the press, the worries are problematic enough to provide some support for considering the less familiar third approach. Further reason for taking the third model seriously draws on moral theoretic considerations.

Chd7 Collaborates with Sox2 to Regulate Activation of Oligodendrocyte Precursor Cells after Spinal Cord Injury.

PubMed

Doi, Toru; Ogata, Toru; Yamauchi, Junji; Sawada, Yasuhiro; Tanaka, Sakae; Nagao, Motoshi

2017-10-25

Oligodendrocyte precursor cells (OPCs) act as a reservoir of new oligodendrocytes (OLs) in homeostatic and pathological conditions. OPCs are activated in response to injury to generate myelinating OLs, but the underlying mechanisms remain poorly understood. Here, we show that chromodomain helicase DNA binding protein 7 (Chd7) regulates OPC activation after spinal cord injury (SCI). Chd7 is expressed in OPCs in the adult spinal cord and its expression is upregulated with a concomitant increase in Sox2 expression after SCI. OPC-specific ablation of Chd7 in injured mice leads to reduced OPC proliferation, the loss of OPC identity, and impaired OPC differentiation. Ablation of Chd7 or Sox2 in cultured OPCs shows similar phenotypes to those observed in Chd7 knock-out mice. Chd7 and Sox2 form a complex in OPCs and bind to the promoters or enhancers of the regulator of cell cycle ( Rgcc ) and protein kinase C θ ( PKC θ) genes, thereby inducing their expression. The expression of Rgcc and PKCθ is reduced in the OPCs of the injured Chd7 knock-out mice. In cultured OPCs, overexpression and knock-down of Rgcc or PKCθ promote and suppress OPC proliferation, respectively. Furthermore, overexpression of both Rgcc and PKCθ rescues the Chd7 deletion phenotypes. Chd7 is thus a key regulator of OPC activation, in which it cooperates with Sox2 and acts via direct induction of Rgcc and PKCθ expression. SIGNIFICANCE STATEMENT Spinal cord injury (SCI) leads to oligodendrocyte (OL) loss and demyelination, along with neuronal death, resulting in impairment of motor or sensory functions. Oligodendrocyte precursor cells (OPCs) activated in response to injury are potential sources of OL replacement and are thought to contribute to remyelination and functional recovery after SCI. However, the molecular mechanisms underlying OPC activation, especially its epigenetic regulation, remain largely unclear. We demonstrate here that the chromatin remodeler chromodomain helicase DNA binding protein 7 (Chd7) regulates the proliferation and identity of OPCs after SCI. We have further identified regulator of cell cycle (Rgcc) and protein kinase Cθ (PKCθ) as novel targets of Chd7 for OPC activation. Copyright © 2017 the authors 0270-6474/17/3710290-20$15.00/0.

Human thrombomodulin knock-in mice reveal differential effects of human thrombomodulin on thrombosis and atherosclerosis.

PubMed

Raife, Thomas J; Dwyre, Denis M; Stevens, Jeff W; Erger, Rochelle A; Leo, Lorie; Wilson, Katina M; Fernández, Jose A; Wilder, Jennifer; Kim, Hyung-Suk; Griffin, John H; Maeda, Nobuyo; Lentz, Steven R

2011-11-01

We sought to develop a murine model to examine the antithrombotic and antiinflammatory functions of human thrombomodulin in vivo. Knock-in mice that express human thrombomodulin from the murine thrombomodulin gene locus were generated. Compared with wild-type mice, human thrombomodulin knock-in mice exhibited decreased protein C activation in the aorta (P<0.01) and lung (P<0.001). Activation of endogenous protein C following infusion of thrombin was decreased by 90% in knock-in mice compared with wild-type mice (P<0.05). Carotid artery thrombosis induced by photochemical injury occurred more rapidly in knock-in mice (12±3 minutes) than in wild-type mice (31±6 minutes; P<0.05). No differences in serum cytokine levels were detected between knock-in and wild-type mice after injection of endotoxin. When crossed with apolipoprotein E-deficient mice and fed a Western diet, knock-in mice had a further decrease in protein C activation but did not exhibit increased atherosclerosis. Expression of human thrombomodulin in place of murine thrombomodulin produces viable mice with a prothrombotic phenotype but unaltered responses to systemic inflammatory or atherogenic stimuli. This humanized animal model will be useful for investigating the function of human thrombomodulin under pathophysiological conditions in vivo.

76 FR 73986 - Redelegation of Authority Under the Age Discrimination Act of 1975

Federal Register 2010, 2011, 2012, 2013, 2014

2011-11-29

... Under the Age Discrimination Act of 1975 AGENCY: Office of the Assistant Secretary for Fair Housing and Equal Opportunity, HUD. ACTION: Notice of redelegation of authority. SUMMARY: The Assistant Secretary for Fair Housing and Equal Opportunity (FHEO) supersedes all prior redelegations of authority made...

7 CFR 3575.40 - Equal opportunity and Fair Housing Act requirements.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 7 Agriculture 15 2010-01-01 2010-01-01 false Equal opportunity and Fair Housing Act requirements. 3575.40 Section 3575.40 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, DEPARTMENT OF AGRICULTURE GENERAL Community Programs Guaranteed Loans § 3575.40 Equal...

Unequal Opportunities: The Recruitment, Selection and Promotion Prospects for Black Teachers.

ERIC Educational Resources Information Center

Brar, Harbhajan Singh

1991-01-01

The 1988 Education Reform Act greatly impacted equal opportunities in recruitment and selection of black teachers in the United Kingdom. A case study examines recruitment and selection in London, noting that ad hoc practices (encouraged by the 1988 Act) perpetuates the poor position of black teachers. (SM)

76 FR 41262 - Notice of Intent To Award Affordable Care Act (ACA) Funding, EH11-1103

Federal Register 2010, 2011, 2012, 2013, 2014

2011-07-13

... Intent To Award Affordable Care Act (ACA) Funding, EH11-1103 Notice of Intent to award Affordable Care... opportunity EH11-1103, ``National Environmental Public Health Tracking Program-Network Implementation... under funding opportunity EH11-1103, ``National Environmental Public Health Tracking Program-Network...

"America COMPETES Reauthorization Act of 2010" or the "America Creating Opportunities to Meaningfully Promote Excellence in Technology, Education, and Science Reauthorization Act of 2010." House of Representatives, One Hundred Eleventh Congress, Second Session. H.R. 5116

ERIC Educational Resources Information Center

US House of Representatives, 2010

2010-01-01

The America COMPETES (Creating Opportunities to Meaningfully Promote Excellence in Technology, Education, and Science) Reauthorization Act of 2010 documented here is divided into the following titles: (1) Office of Science and Technology Policy (Coordination of Federal STEM [science, technology, engineering, and mathematics] education;…

19 CFR 10.841 - Applicability.

Code of Federal Regulations, 2010 CFR

2010-04-01

... Haitian Hemispheric Opportunity through Partnership Encouragement Act of 2006 (HOPE I Act), amended the... Encouragement Act of 2008 (HOPE II Act) amended certain provisions within section 213A. Section 213A of the...

48 CFR 2819.7001 - General.

Code of Federal Regulations, 2010 CFR

2010-10-01

... BUSINESS PROGRAMS Forecasts of Expected Contract Opportunities 2819.7001 General. Section 501 of Public Law 100-656, the Business Opportunity Development Reform Act of 1988, requires executive agencies having... expected contract opportunities, or classes of contract opportunities that small business concerns...

[RS-1 enhanced the efficiency of CRISPR-Cas9 mediated knock-in of human lactoferrin].

PubMed

Zhou, Wenjun; Guo, Rihong; Deng, Mingtian; Wang, Feng; Zhang, Yanli

2017-08-25

This study aims to knock out the goat β-lactoglobulin (BLG) gene using CRISPR-Cas9 system and knock in human lactoferrin (hLF) at the BLG locus, and further study the effect of RAD51 stimulatory compound (RS-1) on homologous recombination efficiency. First, we designed an sgRNA targeting the first exon of goat BLG gene and constructed a co-expression vector pCas9-sgBLG. This sgRNA vector was then transfected into goat ear fibroblasts (GEFs), and the target region was examined by T7EN1 assay and sequencing. Second, we constructed a targeting vector pBHA-hLF-NIE including NEO and EGFP genes based on BLG gene locus. This targeting vector together with pCas9-sgBLG expression vector was co-transfected into GEFs. Transfected cells were then treated with 0, 5, 10 and 20 μmol/L RS-1 for 72 h to analyse the EGFP expression efficiency. Next, we used 800 μg/mL G418 to screen G418-resistent cell clones, and studied hLF site-specific knock-in cell clones by PCR and sequencing. The editing efficiency of sgBLG was between 25% and 31%. The EGFP expression efficiency indicated that the gene knock-in efficiency was improved by RS-1 in a dose-dependent manner, which could reach 3.5-fold compared to the control group. The percentage of positive cells with hLF knock-in was increased to 32.61% when 10 μmol/L RS-1 was used. However, when the concentration of RS-1 increased to 20 μmol/L, the percentage of positive cells decreased to 22.22% and resulted in an increase of senescent cell clone number. These results suggested that hLF knock-in and BLG knock-out in GEFs were achieved by using CRISPR/Cas9 system, and optimum concentration of RS-1 could improve knock-in efficiency, which provides a reference for efficiently obtaining gene knock-in cells using CRISPR/Cas9 in the future.

The Effect of Compression Ratio on Knock Limits of High-Performance Fuels in a CFR Engine II : Blends of 2,2,3-Trimethylpentane with 28-R

NASA Technical Reports Server (NTRS)

Tower, Leonard K

1945-01-01

The knock-limited performance of blends of 0,50; and 100 percent by volume of 2,2,3-trimethylpentane in 28-R fuel determined with a modified F-4 engine at three sets of conditions varying from severe to mild at each of three compression ratios (6.0, 8.0, and 10.0). A comparison of the knock-limited performance of 2,2,3-trimethylpentane with that of triptane (2,2,3-trimethylbutane) is included. The knock-Limited performance of 2,2,3-trimethylpontane was usually more sensitive to either compression ratio or inlet-air temperature than 28-R fuel, but the ratio of the knock-limited indicated mean effective pressure of a given blend containing 2,2,3-trimethypentane and 28-R to the indicated mean effective pressure of 28-R alone was not greatly affected by compression ratio if the engine operating conditions were mild. Although 2,2,3-trimethylpentane in general had a lower knock-limited performance than triptane, the characteristics of the two fuels were somewhat similar.

MMEJ-assisted gene knock-in using TALENs and CRISPR-Cas9 with the PITCh systems.

PubMed

Sakuma, Tetsushi; Nakade, Shota; Sakane, Yuto; Suzuki, Ken-Ichi T; Yamamoto, Takashi

2016-01-01

Programmable nucleases enable engineering of the genome by utilizing endogenous DNA double-strand break (DSB) repair pathways. Although homologous recombination (HR)-mediated gene knock-in is well established, it cannot necessarily be applied in every cell type and organism because of variable HR frequencies. We recently reported an alternative method of gene knock-in, named the PITCh (Precise Integration into Target Chromosome) system, assisted by microhomology-mediated end-joining (MMEJ). MMEJ harnesses independent machinery from HR, and it requires an extremely short homologous sequence (5-25 bp) for DSB repair, resulting in precise gene knock-in with a more easily constructed donor vector. Here we describe a streamlined protocol for PITCh knock-in, including the design and construction of the PITCh vectors, and their delivery to either human cell lines by transfection or to frog embryos by microinjection. The construction of the PITCh vectors requires only a few days, and the entire process takes ∼ 1.5 months to establish knocked-in cells or ∼ 1 week from injection to early genotyping in frog embryos.

Alterations in ethanol-induced behaviors and consumption in knock-in mice expressing ethanol-resistant NMDA receptors.

PubMed

den Hartog, Carolina R; Beckley, Jacob T; Smothers, Thetford C; Lench, Daniel H; Holseberg, Zack L; Fedarovich, Hleb; Gilstrap, Meghin J; Homanics, Gregg E; Woodward, John J

2013-01-01

Ethanol's action on the brain likely reflects altered function of key ion channels such as glutamatergic N-methyl-D-aspartate receptors (NMDARs). In this study, we determined how expression of a mutant GluN1 subunit (F639A) that reduces ethanol inhibition of NMDARs affects ethanol-induced behaviors in mice. Mice homozygous for the F639A allele died prematurely while heterozygous knock-in mice grew and bred normally. Ethanol (44 mM; ∼0.2 g/dl) significantly inhibited NMDA-mediated EPSCs in wild-type mice but had little effect on responses in knock-in mice. Knock-in mice had normal expression of GluN1 and GluN2B protein across different brain regions and a small reduction in levels of GluN2A in medial prefrontal cortex. Ethanol (0.75-2.0 g/kg; i.p.) increased locomotor activity in wild-type mice but had no effect on knock-in mice while MK-801 enhanced activity to the same extent in both groups. Ethanol (2.0 g/kg) reduced rotarod performance equally in both groups but knock-in mice recovered faster following a higher dose (2.5 g/kg). In the elevated zero maze, knock-in mice had a blunted anxiolytic response to ethanol (1.25 g/kg) as compared to wild-type animals. No differences were noted between wild-type and knock-in mice for ethanol-induced loss of righting reflex, sleep time, hypothermia or ethanol metabolism. Knock-in mice consumed less ethanol than wild-type mice during daily limited-access sessions but drank more in an intermittent 24 h access paradigm with no change in taste reactivity or conditioned taste aversion. Overall, these data support the hypothesis that NMDA receptors are important in regulating a specific constellation of effects following exposure to ethanol.

Alterations in Ethanol-Induced Behaviors and Consumption in Knock-In Mice Expressing Ethanol-Resistant NMDA Receptors

PubMed Central

den Hartog, Carolina R.; Beckley, Jacob T.; Smothers, Thetford C.; Lench, Daniel H.; Holseberg, Zack L.; Fedarovich, Hleb; Gilstrap, Meghin J.; Homanics, Gregg E.; Woodward, John J.

2013-01-01

Ethanol's action on the brain likely reflects altered function of key ion channels such as glutamatergic N-methyl-D-aspartate receptors (NMDARs). In this study, we determined how expression of a mutant GluN1 subunit (F639A) that reduces ethanol inhibition of NMDARs affects ethanol-induced behaviors in mice. Mice homozygous for the F639A allele died prematurely while heterozygous knock-in mice grew and bred normally. Ethanol (44 mM; ∼0.2 g/dl) significantly inhibited NMDA-mediated EPSCs in wild-type mice but had little effect on responses in knock-in mice. Knock-in mice had normal expression of GluN1 and GluN2B protein across different brain regions and a small reduction in levels of GluN2A in medial prefrontal cortex. Ethanol (0.75–2.0 g/kg; IP) increased locomotor activity in wild-type mice but had no effect on knock-in mice while MK-801 enhanced activity to the same extent in both groups. Ethanol (2.0 g/kg) reduced rotarod performance equally in both groups but knock-in mice recovered faster following a higher dose (2.5 g/kg). In the elevated zero maze, knock-in mice had a blunted anxiolytic response to ethanol (1.25 g/kg) as compared to wild-type animals. No differences were noted between wild-type and knock-in mice for ethanol-induced loss of righting reflex, sleep time, hypothermia or ethanol metabolism. Knock-in mice consumed less ethanol than wild-type mice during daily limited-access sessions but drank more in an intermittent 24 h access paradigm with no change in taste reactivity or conditioned taste aversion. Overall, these data support the hypothesis that NMDA receptors are important in regulating a specific constellation of effects following exposure to ethanol. PMID:24244696

Heat shock-initiated apoptosis is accelerated and removal of damaged cells is delayed in the testis of clusterin/ApoJ knock-out mice.

PubMed

Bailey, Robert W; Aronow, Bruce; Harmony, Judith A K; Griswold, Michael D

2002-04-01

The secretion and localization of clusterin in the testis has led to the hypothesis that clusterin plays a role in spermatogenesis. Furthermore, the association of clusterin with apoptosis, cellular injury, disease, and regression of nongonadal tissues has led to the hypothesis that clusterin acts to protect cells from apoptosis or may be involved in tissue remodeling. To investigate the role of clusterin in the testis, we analyzed clusterin knock-out (cluKO) mice to determine the impact of the absence of clusterin on spermatogenesis. Furthermore, we investigated the cellular response to injury caused by methoxyacetic acid (MAA) toxicity and mild heat exposure in the cluKO mice to determine the extent to which clusterin protects against apoptosis or participates in tissue remodeling. We found that cluKO mice were fertile and had essentially normal spermatogenesis with the exception of some incomplete spermiation after stage VIII. No differences in testicular morphology or the incidence of apoptosis in the testis were seen between the cluKO and clusterin wild-type (cluWT) mice after MAA treatment. In contrast, apoptosis was delayed in the cluWT mice compared with the cluKO mice after heat exposure, suggesting that clusterin does have a slight protective effect against apoptosis under some conditions. Also, a dramatic loss of germ cells after heat stress occurred earlier in the cluWT testes than in the cluKO testes. Clusterin is clearly acting in a dual role in that cells can be protected from damage and dead cells can be more easily removed after some types of cellular damage but not after others.

Interactions between inflammatory signals and the progesterone receptor in regulating gene expression in pregnant human uterine myocytes

PubMed Central

Lee, Yun; Sooranna, Suren R; Terzidou, Vasso; Christian, Mark; Brosens, Jan; Huhtinen, Kaisa; Poutanen, Matti; Barton, Geraint; Johnson, Mark R; Bennett, Phillip R

2012-01-01

The absence of a fall in circulating progesterone levels has led to the concept that human labour is associated with ‘functional progesterone withdrawal’ caused through changes in the expression or function of progesterone receptor (PR). At the time of labour, the human uterus is heavily infiltrated with inflammatory cells, which release cytokines to create a ‘myometrial inflammation’ via NF-κB activation. The negative interaction between NF-κB and PR, may represent a mechanism to account for ‘functional progesterone withdrawal’ at term. Conversely, PR may act to inhibit NF-κB function and so play a role in inhibition of myometrial inflammation during pregnancy. To model this inter-relationship, we have used small interfering (si) RNA-mediated knock-down of PR in human pregnant myocytes and whole genome microarray analysis to identify genes regulated through PR. We then activated myometrial inflammation using IL-1β stimulation to determine the role of PR in myometrial inflammation regulation. Through PR-knock-down, we found that PR regulates gene networks involved in myometrial quiescence and extracellular matrix integrity. Activation of myometrial inflammation was found to antagonize PR-induced gene expression, of genes normally upregulated via PR. We found that PR does not play a role in repression of pro-inflammatory gene networks induced by IL-1β and that only MMP10 was significantly regulated in opposite directions by IL-1β and PR. We conclude that progesterone acting through PR does not generally inhibit myometrial inflammation. Activation of myometrial inflammation does cause ‘functional progesterone withdrawal’ but only in the context of genes normally upregulated via PR. PMID:22435466

Disruption of the GDP-mannose synthesis pathway in Streptomyces coelicolor results in antibiotic hyper-susceptible phenotypes.

PubMed

Howlett, Robert; Anttonen, Katri; Read, Nicholas; Smith, Margaret C M

2018-04-01

Actinomycete bacteria use polyprenol phosphate mannose as a lipid linked sugar donor for extra-cytoplasmic glycosyl transferases that transfer mannose to cell envelope polymers, including glycoproteins and glycolipids. We showed recently that strains of Streptomyces coelicolor with mutations in the gene ppm1 encoding polyprenol phosphate mannose synthase were both resistant to phage φC31 and have greatly increased susceptibility to antibiotics that mostly act on cell wall biogenesis. Here we show that mutations in the genes encoding enzymes that act upstream of Ppm1 in the polyprenol phosphate mannose synthesis pathway can also confer phage resistance and antibiotic hyper-susceptibility. GDP-mannose is a substrate for Ppm1 and is synthesised by GDP-mannose pyrophosphorylase (GMP; ManC) which uses GTP and mannose-1-phosphate as substrates. Phosphomannomutase (PMM; ManB) converts mannose-6-phosphate to mannose-1-phosphate. S. coelicolor strains with knocked down GMP activity or with a mutation in sco3028 encoding PMM acquire phenotypes that resemble those of the ppm1 - mutants i.e. φC31 resistant and susceptible to antibiotics. Differences in the phenotypes of the strains were observed, however. While the ppm1 - strains have a small colony phenotype, the sco3028 :: Tn5062 mutants had an extremely small colony phenotype indicative of an even greater growth defect. Moreover we were unable to generate a strain in which GMP activity encoded by sco3039 and sco4238 is completely knocked out, indicating that GMP is also an important enzyme for growth. Possibly GDP-mannose is at a metabolic branch point that supplies alternative nucleotide sugar donors.

Health Careers Opportunity Program (HCOP). Section 787 (Public Health Service Act). Program Guide.

ERIC Educational Resources Information Center

Department of Health and Human Services, Washington, DC.

This document summarizes the requirements and guidelines for the Health Careers Opportunity Program (HCOP). This program is authorized by Section 787 of the Public Health Service Act to make grants to and contracts with postsecondary institutions to carry out programs which assist individuals from disadvantaged backgrounds to enter and graduate…

29 CFR 1614.703 - Manner and format of data.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 29 Labor 4 2010-07-01 2010-07-01 false Manner and format of data. 1614.703 Section 1614.703 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION FEDERAL SECTOR EQUAL EMPLOYMENT OPPORTUNITY Procedures Under the Notification and Federal Employee Antidiscrimination and Retaliation Act of 2002 (No FEAR Act) §...

Royal Wine Corporation d/b/a/ Royal Kedem (Herzog Wine Cellars), Oxnard, CA; Proposed Settlement of Clean Water Act Class II Administrative Penalty and Opportunity to Comment

EPA Pesticide Factsheets

Public Notice of Proposed Settlement of Clean Water Act Class II Administrative Penalty and Opportunity to Comment In the Matter of Royal Wine Corporation d/b/a/ Royal Kedem (Herzog Wine Cellars), Oxnard, California.

78 FR 42103 - Agency Information Collection Activities: African Growth and Opportunity Act Certificate of Origin

Federal Register 2010, 2011, 2012, 2013, 2014

2013-07-15

... Activities: African Growth and Opportunity Act Certificate of Origin AGENCY: U.S. Customs and Border... submitting the following information collection request to the Office of Management and Budget (OMB) for... Certificate of Origin (AGOA). This is a proposed extension of an information collection that was previously...

TOPS Report: Analysis of the TOPS Program from 2006-2015

ERIC Educational Resources Information Center

Louisiana Board of Regents, 2016

2016-01-01

The Tuition Opportunity Program for Students (subsequently renamed the Taylor Opportunity Program for Students, or TOPS) was created via ACT 1375 during the 1997 Regular Legislative Session. The first college freshman class to receive TOPS awards entered postsecondary education in the fall of 1998. ACT 1202 of the 2001 Regular Legislative Session…

77 FR 33223 - Announcement of the Publication of Funding Opportunity Announcements Under the Runaway and...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-06-05

... the Publication of Funding Opportunity Announcements Under the Runaway and Homeless Youth Act AGENCY... Statutory Authority: Runaway and Homeless Youth Act, 42 U.S.C. sections 5701-5752, as amended by the.... Porter, Director, Runaway and Homeless Youth Program, Family and Youth Services Bureau, 1250 Maryland Ave...

76 FR 54774 - Notice of Intent To Award Affordable Care Act Funding, Funding Opportunity Announcement CDC-RFA...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-09-02

... Intent To Award Affordable Care Act Funding, Funding Opportunity Announcement CDC-RFA-DP09-905 AGENCY: Centers for Disease Control and Prevention (CDC), Department of Health and Human Services (HHS). ACTION: Notice. SUMMARY: This notice provides notice of CDC's intent to fund continuation cooperative agreement...

29 CFR 37.10 - To what extent are employment practices covered by this part?

Code of Federal Regulations, 2010 CFR

2010-07-01

... AND EQUAL OPPORTUNITY PROVISIONS OF THE WORKFORCE INVESTMENT ACT OF 1998 (WIA) General Provisions § 37... Opportunity Commission (EEOC) regulations, guidance and appropriate case law in determining whether a... Immigration and Nationality Act should be aware of the obligations imposed by that provision. See 8 U.S.C...

29 CFR 1614.703 - Manner and format of data.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 29 Labor 4 2012-07-01 2012-07-01 false Manner and format of data. 1614.703 Section 1614.703 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION FEDERAL SECTOR EQUAL EMPLOYMENT OPPORTUNITY Procedures Under the Notification and Federal Employee Antidiscrimination and Retaliation Act of 2002 (No FEAR Act) §...

29 CFR 1614.703 - Manner and format of data.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 29 Labor 4 2014-07-01 2014-07-01 false Manner and format of data. 1614.703 Section 1614.703 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION FEDERAL SECTOR EQUAL EMPLOYMENT OPPORTUNITY Procedures Under the Notification and Federal Employee Antidiscrimination and Retaliation Act of 2002 (No FEAR Act) §...

29 CFR 1614.703 - Manner and format of data.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 29 Labor 4 2013-07-01 2013-07-01 false Manner and format of data. 1614.703 Section 1614.703 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION FEDERAL SECTOR EQUAL EMPLOYMENT OPPORTUNITY Procedures Under the Notification and Federal Employee Antidiscrimination and Retaliation Act of 2002 (No FEAR Act) §...

Culture time of vitrified/warmed zygotes before microinjection affects the production efficiency of CRISPR-Cas9-mediated knock-in mice.

PubMed

Nakagawa, Yoshiko; Sakuma, Tetsushi; Nishimichi, Norihisa; Yokosaki, Yasuyuki; Takeo, Toru; Nakagata, Naomi; Yamamoto, Takashi

2017-05-15

Robust reproductive engineering techniques are required for the efficient and rapid production of genetically modified mice. We have reported the efficient production of genome-edited mice using reproductive engineering techniques, such as ultra-superovulation, in vitro fertilization (IVF) and vitrification/warming of zygotes. We usually use vitrified/warmed fertilized oocytes created by IVF for microinjection because of work efficiency and flexible scheduling. Here, we investigated whether the culture time of zygotes before microinjection influences the efficiency of producing knock-in mice. Knock-in mice were generated using clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9) system and single-stranded oligodeoxynucleotide (ssODN) or PITCh (Precise Integration into Target Chromosome) system, a method of integrating a donor vector assisted by microhomology-mediated end-joining. The cryopreserved fertilized oocytes were warmed, cultured for several hours and microinjected at different timings. Microinjection was performed with Cas9 protein, guide RNA(s), and an ssODN or PITCh donor plasmid for the ssODN knock-in and the PITCh knock-in, respectively. Different production efficiencies of knock-in mice were observed by changing the timing of microinjection. Our study provides useful information for the CRISPR-Cas9-based generation of knock-in mice. © 2017. Published by The Company of Biologists Ltd.

Highly efficient CRISPR/HDR-mediated knock-in for mouse embryonic stem cells and zygotes.

PubMed

Wang, Bangmei; Li, Kunyu; Wang, Amy; Reiser, Michelle; Saunders, Thom; Lockey, Richard F; Wang, Jia-Wang

2015-10-01

The clustered regularly interspaced short palindromic repeat (CRISPR) gene editing technique, based on the non-homologous end-joining (NHEJ) repair pathway, has been used to generate gene knock-outs with variable sizes of small insertion/deletions with high efficiency. More precise genome editing, either the insertion or deletion of a desired fragment, can be done by combining the homology-directed-repair (HDR) pathway with CRISPR cleavage. However, HDR-mediated gene knock-in experiments are typically inefficient, and there have been no reports of successful gene knock-in with DNA fragments larger than 4 kb. Here, we describe the targeted insertion of large DNA fragments (7.4 and 5.8 kb) into the genomes of mouse embryonic stem (ES) cells and zygotes, respectively, using the CRISPR/HDR technique without NHEJ inhibitors. Our data show that CRISPR/HDR without NHEJ inhibitors can result in highly efficient gene knock-in, equivalent to CRISPR/HDR with NHEJ inhibitors. Although NHEJ is the dominant repair pathway associated with CRISPR-mediated double-strand breaks (DSBs), and biallelic gene knock-ins are common, NHEJ and biallelic gene knock-ins were not detected. Our results demonstrate that efficient targeted insertion of large DNA fragments without NHEJ inhibitors is possible, a result that should stimulate interest in understanding the mechanisms of high efficiency CRISPR targeting in general.

Motor Deficits and Cerebellar Atrophy in Elovl5 Knock Out Mice.

PubMed

Hoxha, Eriola; Gabriele, Rebecca M C; Balbo, Ilaria; Ravera, Francesco; Masante, Linda; Zambelli, Vanessa; Albergo, Cristian; Mitro, Nico; Caruso, Donatella; Di Gregorio, Eleonora; Brusco, Alfredo; Borroni, Barbara; Tempia, Filippo

2017-01-01

Spino-Cerebellar-Ataxia type 38 (SCA38) is caused by missense mutations in the very long chain fatty acid elongase 5 gene, ELOVL5 . The main clinical findings in this disease are ataxia, hyposmia and cerebellar atrophy. Mice in which Elovl5 has been knocked out represent a model of the loss of function hypothesis of SCA38. In agreement with this hypothesis, Elovl5 knock out mice reproduced the main symptoms of patients, motor deficits at the beam balance test and hyposmia. The cerebellar cortex of Elovl5 knock out mice showed a reduction of thickness of the molecular layer, already detectable at 6 months of age, confirmed at 12 and 18 months. The total perimeter length of the Purkinje cell (PC) layer was also reduced in Elovl5 knock out mice. Since Elovl5 transcripts are expressed by PCs, whose dendrites are a major component of the molecular layer, we hypothesized that an alteration of their dendrites might be responsible for the reduced thickness of this layer. Reconstruction of the dendritic tree of biocytin-filled PCs, followed by Sholl analysis, showed that the distribution of distal dendrites was significantly reduced in Elovl5 knock out mice. Dendritic spine density was conserved. These results suggest that Elovl5 knock out mice recapitulate SCA38 symptoms and that their cerebellar atrophy is due, at least in part, to a reduced extension of PC dendritic arborization.

Strong morphological defects in conditional Arabidopsis abp1 knock-down mutants generated in absence of functional ABP1 protein.

PubMed

Michalko, Jaroslav; Glanc, Matouš; Perrot-Rechenmann, Catherine; Friml, Jiří

2016-01-01

The Auxin Binding Protein 1 (ABP1) is one of the most studied proteins in plants. Since decades ago, it has been the prime receptor candidate for the plant hormone auxin with a plethora of described functions in auxin signaling and development. The developmental importance of ABP1 has recently been questioned by identification of Arabidopsis thaliana abp1 knock-out alleles that show no obvious phenotypes under normal growth conditions. In this study, we examined the contradiction between the normal growth and development of the abp1 knock-outs and the strong morphological defects observed in three different ethanol-inducible abp1 knock-down mutants ( abp1-AS, SS12K, SS12S). By analyzing segregating populations of abp1 knock-out vs. abp1 knock-down crosses we show that the strong morphological defects that were believed to be the result of conditional down-regulation of ABP1 can be reproduced also in the absence of the functional ABP1 protein. This data suggests that the phenotypes in  abp1 knock-down lines are due to the off-target effects and asks for further reflections on the biological function of ABP1 or alternative explanations for the missing phenotypic defects in the abp1 loss-of-function alleles.

[Establishment of L-periaxin gene knock-out RSC96 cell line].

PubMed

Liang, Min; Peng, Tingting; Shi, Yawei

2016-12-25

Periaxin, a protein of noncompact myelin, is specifically expressed in the peripheral nervous system (PNS). There are two protein isoform L-periaxin and S-Periaxin by alternative splicing of periaxin gene, playing an important role in the initiation of myelin formation. So far, 18 different mutation sites in L-periaxin gene have been found to induce the peripheral demyelinating neurological charcot-marie-tooth diseases subtype 4F (CMT4F). The technique of activation of transcription activator-like effector nucleases (TALENS) was used to knock out the L-periaxin gene in RSC 96 cell line of Rattus. According to the design principle, the knock-out site of L-periaxin was assured to NLS domain of L-periaxin, which is target sequence of left and right arms of TALEN. The knock-out vectors of TALEN-L and TALEN-R were established and transfected into RSC96 cell. After puromycin screening, L-periaxin was knocked out successfully in RSC96 cell, which is confirmed by DNA sequence. The mutation efficiency is 21.6%. S-periaxin, not L-periaxin can be detected by Western blotting in L-periaxin gene knock-out RSC96 cell. The cell growth rate was decreased and the number of cells in G1 increased and decreased in S phase in L-periaxin gene knock-out RSC96 cell by flow cytometry and MTT assay.

Measurement of Knock Characteristics in Spark-ignition Engines

NASA Technical Reports Server (NTRS)

Schutz, R

1940-01-01

This paper presents a discussion of three potential sources of error in recording engine knocking which are: the natural oscillation of the membrane, the shock process between test contacts, and the danger of burned contacts. Following this discussion, the paper calls attention to various results which make the bouncing-pin indicator appear fundamentally unsuitable for recording knock phenomena.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yang, Yi; Dec, John E.; Sjoberg, Magnus

Modern spark-ignition (SI) engine technologies have considerably changed in-cylinder conditions under which fuel autoignition and engine knock take place. In this paper, fundamental HCCI engine experiments are proposed as a means for characterizing the impact of these technologies on the knock propensity of different fuels. In particular, the impacts of turbocharging, direct injection (DI), and downspeeding on operation with ethanol and gasoline are investigated to demonstrate this approach. Results reported earlier for ethanol and gasoline on HCCI combustion are revisited with the new perspective of how their autoignition characteristics fit into the anti-knock requirement in modern SI engines. For example,more » the weak sensitivity to pressure boost demonstrated by ethanol in HCCI autoignition can be used to explain the strong knock resistance of ethanol fuels for turbocharged SI engines. Further, ethanol's high sensitivity to charge temperature makes charge cooling, which can be produced by fuel vaporization via direct injection or by piston expansion via spark-timing retard, very effective for inhibiting knock. On the other hand, gasoline autoignition shows a higher sensitivity to pressure, so only very low pressure boost can be applied before knock occurs. Gasoline also demonstrates low temperature sensitivity, so it is unable to make as effective use of the charge cooling produced by fuel vaporization or spark retard. These arguments comprehensively explain literature results on ethanol's substantially better anti-knock performance over gasoline in modern turbocharged DISI engines. Fundamental HCCI experiments such as these can thus be used as a diagnostic and predictive tool for knock-limited SI engine performance for various fuels. As a result, examples are presented where HCCI experiments are used to identify biofuel compounds with good potential for modern SI-engine applications.« less

Electromyographic evidence in support of a knock-in mouse model of DYT1 Dystonia.

PubMed

DeAndrade, Mark P; Trongnetrpunya, Amy; Yokoi, Fumiaki; Cheetham, Chad C; Peng, Ning; Wyss, J Michael; Ding, Mingzhou; Li, Yuqing

2016-11-01

DYT1 dystonia is an autosomal-dominant movement disorder characterized by abnormal, often repetitive, movements and postures. Its hallmark feature is sustained or intermittent contractions of muscles involving co-contractions of antagonist muscle pairs. The symptoms are relieved with the anticholinergic drug trihexyphenidyl. The primary mutation is a trinucleotide deletion (ΔGAG) in DYT1/TOR1A, which codes for torsinA. Previous studies showed that (1) heterozygous Dyt1 ΔGAG knock-in mice, which have an analogous mutation in the endogenous gene, exhibit motor deficits and altered corticostriatal synaptic plasticity in the brain and (2) these deficits can be rescued by trihexyphenidyl. However, brain imaging studies suggest that the Dyt1 knock-in mouse models nonmanifesting mutation carriers of DYT1 dystonia. The aim of this work was to examine the hallmark features of DYT1 dystonia in the Dyt1 knock-in mice by analyzing muscular activities. Wireless telemetry devices with biopotential channels were implanted to the bicep and the rectus femori muscles in Dyt1 knock-in mice, and muscular activities were recorded before and after trihexyphenidyl administration. (1) Consistent with DYT1 dystonia patients, Dyt1 knock-in mice showed sustained contractions and co-contractions of the antagonistic bicep femoris and rectus femoris. (2) The abnormal muscle contractions were normalized by trihexyphenidyl. The results suggest that the motor deficits in Dyt1 knock-in mice are likely produced by abnormal muscle contractions, and Dyt1 knock-in mice can potentially be used as a manifesting disease model to study pathophysiology and develop novel therapeutics. © 2016 International Parkinson and Movement Disorder Society. © 2016 International Parkinson and Movement Disorder Society.

Signal Transducer and Activator of Transcription 1 (STAT1) Knock-down Induces Apoptosis in Malignant Pleural Mesothelioma.

PubMed

Arzt, Lisa; Halbwedl, Iris; Gogg-Kamerer, Margit; Popper, Helmut H

2017-07-01

Malignant pleural mesothelioma (MPM) is the most common primary tumor of the pleura. Its incidence is still increasing in Europe and the prognosis remains poor. We investigated the oncogenic function of signal transducer and activator of transcription 1 (STAT1) in MPM in more detail. A miRNA profiling was performed on 52 MPM tissue samples. Upregulated miRNAs (targeting SOCS1/3) were knocked-down using miRNA inhibitors. mRNA expression levels of STAT1/3, SOCS1/3 were detected in MPM cell lines. STAT1 has been knocked-down using siRNA and qPCR was used to detect mRNA expression levels of all JAK/STAT family members and genes that regulate them. An immunohistochemical staining was performed to detect the expression of caspases. STAT1 was upregulated and STAT3 was downregulated, SOCS1/3 protein was not detected but it was possible to detect SOCS1/3 mRNA in MPM cell lines. The upregulated miRNAs were successfully knocked-down, however the expected effect on SOCS1 expression was not detected. STAT1 knock-down had different effects on STAT3/5 expression. Caspase 3a and 8 expression was found to be increased after STAT1 knock-down. The physiologic regulation of STAT1 via SOCS1 is completely lost in MPM and it does not seem that the miRNAs identified by now, do inhibit the expression of SOCS1. MPM cell lines compensate STAT1 knock-down by increasing the expression of STAT3 or STAT5a, two genes which are generally considered to be oncogenes. And much more important, STAT1 knock-down induces apoptosis in MPM cell lines and STAT1 might therefore be a target for therapeutic intervention.

Knock-in fibroblasts and transgenic blastocysts for expression of human FGF2 in the bovine β-casein gene locus using CRISPR/Cas9 nuclease-mediated homologous recombination.

PubMed

Jeong, Young-Hee; Kim, Yeong Ji; Kim, Eun Young; Kim, Se Eun; Kim, Jiwoo; Park, Min Jee; Lee, Hong-Gu; Park, Se Pill; Kang, Man-Jong

2016-06-01

Many transgenic domestic animals have been developed to produce therapeutic proteins in the mammary gland, and this approach is one of the most important methods for agricultural and biomedical applications. However, expression and secretion of a protein varies because transgenes are integrated at random sites in the genome. In addition, distal enhancers are very important for transcriptional gene regulation and tissue-specific gene expression. Development of a vector system regulated accurately in the genome is needed to improve production of therapeutic proteins. The objective of this study was to develop a knock-in system for expression of human fibroblast growth factor 2 (FGF2) in the bovine β-casein gene locus. The F2A sequence was fused to the human FGF2 gene and inserted into exon 3 of the β-casein gene. We detected expression of human FGF2 mRNA in the HC11 mouse mammary epithelial cells by RT-PCR and human FGF2 protein in the culture media using western blot analysis when the knock-in vector was introduced. We transfected the knock-in vector into bovine ear fibroblasts and produced knock-in fibroblasts using the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system. Moreover, the CRISPR/Cas9 system was more efficient than conventional methods. In addition, we produced knock-in blastocysts by somatic cell nuclear transfer using the knock-in fibroblasts. Our knock-in fibroblasts may help to create cloned embryos for development of transgenic dairy cattle expressing human FGF2 protein in the mammary gland via the expression system of the bovine β-casein gene.

The Interpretation and Effective Implementation of the Stevenson-Wydler Technology Innovation Act of 1996

NASA Technical Reports Server (NTRS)

Heard, Pamala D.

1996-01-01

The purpose of this research is to develop a plan that gives schools the opportunity to acquire knowledge of the Stevenson-Wydler Technology Innovation Act. The law suggests that federal agencies give surplus equipment to educational entities. It would then become the school's choice to take advantage of the opportunity. The purpose of this proposal is to develop an ongoing partnership with schools around the United States. Its attempt is to inform schools about the law and organize a plan that will allow schools to take advantage of this opportunity. It would inform teachers about available resources and expose their students to educational opportunities.

[Application of atomic absorption spectrometry in the engine knock detection].

PubMed

Chen, Li-Dan

2013-02-01

Because existing human experience diagnosis method and apparatus for auxiliary diagnosis method are difficult to diagnose quickly engine knock. Atomic absorption spectrometry was used to detect the automobile engine knock in in innovative way. After having determined Fe, Al, Cu, Cr and Pb content in the 35 groups of Audi A6 engine oil whose travel course is 2 000 -70 000 kilometers and whose sampling interval is 2 000 kilometers by atomic absorption spectrometry, the database of primary metal content in the same automobile engine at different mileage was established. The research shows that the main metal content fluctuates within a certain range. In practical engineering applications, after the determination of engine oil main metal content and comparison with its database value, it can not only help to diagnose the type and location of engine knock without the disintegration and reduce vehicle maintenance costs and improve the accuracy of engine knock fault diagnosis.

Elevated blood pressure, heart rate and body temperature in mice lacking the XLαs protein of the Gnas locus is due to increased sympathetic tone

PubMed Central

Nunn, Nicolas; Feetham, Claire H; Martin, Jennifer; Barrett-Jolley, Richard; Plagge, Antonius

2013-01-01

New Findings What is the central question of this study? Previously, we showed that Gnasxl knock-out mice are lean and hypermetabolic, with increased sympathetic stimulation of adipose tissue. Do these mice also display elevated sympathetic cardiovascular tone? Is the brain glucagon-like peptide-1 system involved? What is the main finding and its importance? Gnasxl knock-outs have increased blood pressure, heart rate and body temperature. Heart rate variability analysis suggests an elevated sympathetic tone. The sympatholytic reserpine had stronger effects on blood pressure, heart rate and heart rate variability in knock-out compared with wild-type mice. Stimulation of the glucagon-like peptide-1 system inhibited parasympathetic tone to a similar extent in both genotypes, with a stronger associated increase in heart rate in knock-outs. Deficiency of Gnasxl increases sympathetic cardiovascular tone. Imbalances of energy homeostasis are often associated with cardiovascular complications. Previous work has shown that Gnasxl-deficient mice have a lean and hypermetabolic phenotype, with increased sympathetic stimulation of adipose tissue. The Gnasxl transcript from the imprinted Gnas locus encodes the trimeric G-protein subunit XLαs, which is expressed in brain regions that regulate energy homeostasis and sympathetic nervous system (SNS) activity. To determine whether Gnasxl knock-out (KO) mice display additional SNS-related phenotypes, we have now investigated the cardiovascular system. The Gnasxl KO mice were ∼20 mmHg hypertensive in comparison to wild-type (WT) littermates (P≤ 0.05) and hypersensitive to the sympatholytic drug reserpine. Using telemetry, we detected an increased waking heart rate in conscious KOs (630 ± 10 versus 584 ± 12 beats min−1, KO versus WT, P≤ 0.05). Body temperature was also elevated (38.1 ± 0.3 versus 36.9 ± 0.4°C, KO versus WT, P≤ 0.05). To investigate autonomic nervous system influences, we used heart rate variability analyses. We empirically defined frequency power bands using atropine and reserpine and verified high-frequency (HF) power and low-frequency (LF) LF/HF power ratio to be indicators of parasympathetic and sympathetic activity, respectively. The LF/HF power ratio was greater in KOs and more sensitive to reserpine than in WTs, consistent with elevated SNS activity. In contrast, atropine and exendin-4, a centrally acting agonist of the glucagon-like peptide-1 receptor, which influences cardiovascular physiology and metabolism, reduced HF power equally in both genotypes. This was associated with a greater increase in heart rate in KOs. Mild stress had a blunted effect on the LF/HF ratio in KOs consistent with elevated basal sympathetic activity. We conclude that XLαs is required for the inhibition of sympathetic outflow towards cardiovascular and metabolically relevant tissues. PMID:23748904

12 CFR 232.1 - Scope, General Prohibition and Definitions

Code of Federal Regulations, 2010 CFR

2010-01-01

... eligibility, for credit, except as provided in this section. (c) Definitions. (1) Consumer means an individual. (2) Credit has the same meaning as in section 702 of the Equal Credit Opportunity Act, 15 U.S.C. 1691a. (3) Creditor has the same meaning as in section 702 of the Equal Credit Opportunity Act, 15 U.S.C...

ServiceNation: A Call to Higher Education

ERIC Educational Resources Information Center

Jacoby, Barbara

2009-01-01

The Serve America Act was signed into law in April 2009. The Act provides for many opportunities to serve and learn for pre-college youth, college students and graduates, and even baby boomers. It dramatically increases intensive service opportunities by setting AmeriCorps on a path from 75,000 positions annually to 250,000 by 2017, focusing that…

76 FR 21906 - Notice of Intent To Award Affordable Care Act Funding to Approved Applications Formerly Received...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-04-19

... Intent To Award Affordable Care Act Funding to Approved Applications Formerly Received in Response to the Centers for Disease Control and Prevention Funding Opportunity IP11-010, ``Enhanced Surveillance for New... response to CDC Funding Opportunity, CDC-RFA-IP11-010, ``Enhanced Surveillance for New Vaccine Preventable...

3 CFR 8468 - Proclamation 8468 of December 23, 2009. To Take Certain Actions Under the African Growth and...

Code of Federal Regulations, 2010 CFR

2010-01-01

... Certain Actions Under the African Growth and Opportunity Act 8468 Proclamation 8468 Presidential Documents Proclamations Proclamation 8468 of December 23, 2009 Proc. 8468 To Take Certain Actions Under the African Growth... section 111(a) of the African Growth and Opportunity Act (title I of Public Law 106-200) (AGOA...

Evaluation of the DC Opportunity Scholarship Program: First Year Report on Participation

ERIC Educational Resources Information Center

Wolf, Patrick; Eissa, Nada; Puma, Michael

2005-01-01

The District of Columbia School Choice Incentive Act of 2003 was passed by Congress in January 2004. The Act provided funds for District of Columbia Public Schools (DCPS) improvement activities and charter school facility acquisitions. Most notably, the statute established what is now called the DC Opportunity Scholarship Program?the first federal…

Education Issues Raised by S.744: The Border Security, Economic Opportunity, and Immigration Modernization Act

ERIC Educational Resources Information Center

Murphy, Garrett

2014-01-01

This brief report summarizes the requirements for undocumented immigrants set forth by the Border Security, Economic Opportunity, and Immigration Modernization Act (S.744). Assuming that S.744 will move forward in Congress, the report also examines issues having to do with certain language, civics and government, and education/training provisions…

Using RNA interference to knock down the adhesion protein TES.

PubMed

Griffith, Elen

2007-01-01

RNA interference (RNAi) is a specific and efficient method to knock down protein levels using small interfering RNAs (siRNAs), which target mRNA degradation. RNAi can be used in mammalian cell culture systems to target any protein of interest, and several studies have used this method to knock down adhesion proteins. We used siRNAs to knock down the levels of TES, a focal adhesion protein, in HeLa cells. We demonstrated knockdown of both TES mRNA and TES protein. Although total knockdown of TES was not achieved, the observed reduction in TES protein was sufficient to result in a cellular phenotype of reduced actin stress fibers.

The Knock-Limited Performance of Fuel Blends Containing Spiropentane, Methylenecyclobutane, Di-Tert-Butyl Ether, Methyl Tert-Butyl Ether, and Triptane

NASA Technical Reports Server (NTRS)

Meyer, Carl L.

1946-01-01

Tests show that at inlet-air temperatures of 250 deg F and 100 deg F the knock-limited performance of the base fuel of blends, leaded with 4 ml TEL per gallon and containing 20 percent spiropentane, was reduced at fuel/air ratios below 0.085. The 20 percent methylenecyclobutane reduced the knock-limited power of the base fuel at fuel/air ratios below 0.112. Di-tert-butyl ether, methyl-tert-butyl ether, and triptane increased the knock-limited power of the base fuel at all fuel/air ratios and at both temperatures.

Analysis of Spark-Ignition Engine Knock as Seen in Photographs Taken at 200,000 Frames Per Second

NASA Technical Reports Server (NTRS)

Miller, Cearcy D; Olsen, H Lowell; Logan, Walter O , Jr; Osterstrom, Gordon E

1946-01-01

A motion picture of the development of knock in a spark-ignition engine is presented, which consists of 20 photographs taken at intervals of 5 microseconds, or at a rate of 200,000 photographs per second, with an equivalent wide-open exposure time of 6.4 microseconds for each photograph. A motion picture of a complete combustion process, including the development of knock, taken at the rate of 40,000 photographs per second is also presented to assist the reader in orienting the photographs of the knock development taken at 200,000 frames per second.

ACTS of Education

NASA Technical Reports Server (NTRS)

Bauer, Robert; Krawczyk, Richard; Gargione, Frank; Kruse, Hans; Vrotsos, Pete (Technical Monitor)

2002-01-01

Now in its ninth year of operations, the Advanced Communications Technology Satellite (ACTS) program has continued, although since May 2000 in a new operations arrangement involving a university based consortium, the Ohio Consortium for Advanced Communications Technology (OCACT), While NASA has concluded its experimental intentions of ACTS, the spacecraft's ongoing viability has permitted its further operations to provide educational opportunities to engineering and communications students interested in satellite operations, as well as a Ka-band test bed for commercial interests in utilizing Kaband space communications. The consortium has reached its first year of operations. This generous opportunity by NASA has already resulted in unique educational opportunities for students in obtaining "hands-on" experience, such as, in satellite attitude control. An update is presented on the spacecraft and consortium operations.

Efficient generation of knock-in transgenic zebrafish carrying reporter/driver genes by CRISPR/Cas9-mediated genome engineering.

PubMed

Kimura, Yukiko; Hisano, Yu; Kawahara, Atsuo; Higashijima, Shin-ichi

2014-10-08

The type II bacterial CRISPR/Cas9 system is rapidly becoming popular for genome-engineering due to its simplicity, flexibility, and high efficiency. Recently, targeted knock-in of a long DNA fragment via homology-independent DNA repair has been achieved in zebrafish using CRISPR/Cas9 system. This raised the possibility that knock-in transgenic zebrafish could be efficiently generated using CRISPR/Cas9. However, how widely this method can be applied for the targeting integration of foreign genes into endogenous genomic loci is unclear. Here, we report efficient generation of knock-in transgenic zebrafish that have cell-type specific Gal4 or reporter gene expression. A donor plasmid containing a heat-shock promoter was co-injected with a short guide RNA (sgRNA) targeted for genome digestion, a sgRNA targeted for donor plasmid digestion, and Cas9 mRNA. We have succeeded in establishing stable knock-in transgenic fish with several different constructs for 4 genetic loci at a frequency being exceeding 25%. Due to its simplicity, design flexibility, and high efficiency, we propose that CRISPR/Cas9-mediated knock-in will become a standard method for the generation transgenic zebrafish.

Closed loop statistical performance analysis of N-K knock controllers

NASA Astrophysics Data System (ADS)

Peyton Jones, James C.; Shayestehmanesh, Saeed; Frey, Jesse

2017-09-01

The closed loop performance of engine knock controllers cannot be rigorously assessed from single experiments or simulations because knock behaves as a random process and therefore the response belongs to a random distribution also. In this work a new method is proposed for computing the distributions and expected values of the closed loop response, both in steady state and in response to disturbances. The method takes as its input the control law, and the knock propensity characteristic of the engine which is mapped from open loop steady state tests. The method is applicable to the 'n-k' class of knock controllers in which the control action is a function only of the number of cycles n since the last control move, and the number k of knock events that have occurred in this time. A Cumulative Summation (CumSum) based controller falls within this category, and the method is used to investigate the performance of the controller in a deeper and more rigorous way than has previously been possible. The results are validated using onerous Monte Carlo simulations, which confirm both the validity of the method and its high computational efficiency.

Optimization of the production of knock-in alleles by CRISPR/Cas9 microinjection into the mouse zygote.

PubMed

Raveux, Aurélien; Vandormael-Pournin, Sandrine; Cohen-Tannoudji, Michel

2017-02-17

Microinjection of the CRISPR/Cas9 system in zygotes is an efficient and comparatively fast method to generate genetically modified mice. So far, only few knock-in mice have been generated using this approach, and because no systematic study has been performed, parameters controlling the efficacy of CRISPR/Cas9-mediated targeted insertion are not fully established. Here, we evaluated the effect of several parameters on knock-in efficiency changing only one variable at a time. We found that knock-in efficiency was dependent on injected Cas9 mRNA and single-guide RNA concentrations and that cytoplasmic injection resulted in more genotypic complexity compared to pronuclear injection. Our results also indicated that injection into the pronucleus compared to the cytoplasm is preferable to generate knock-in alleles with an oligonucleotide or a circular plasmid. Finally, we showed that Cas9D10A nickase variant was less efficient than wild-type Cas9 for generating knock-in alleles and caused a higher rate of mosaicism. Thus, our study provides valuable information that will help to improve the future production of precise genetic modifications in mice.

Enhanced serotonin response in the hippocampus of Galphaz protein knock-out mice.

PubMed

Oleskevich, Sharon; Leck, Kwong-Joo; Matthaei, Klaus; Hendry, Ian A

2005-06-21

The serotonin-1A [5-hydroxytryptamine 1A (5HT1A)] receptor is important for emotional and homeostatic processes in the central nervous system. In the hippocampus, the 5HT1A receptor couples to inhibitory Gi/o proteins to decrease pyramidal cell excitability. Here we investigate the 5HT1A receptor in a mouse deficient in the alpha-subunit of Gz protein (Galphaz knock-out). Behavioural tests showed heightened anxiety and depression-like behaviour in the Galphaz knock-out mice. Whole-cell recording in CA1 pyramidal neurons showed a significantly greater 5HT1A receptor-mediated potassium current in Galphaz knock-out mice. The effect was independent of 5HT4 receptors as the slow after-hyperpolarization was unaffected and a slow depolarization was absent in the Galphaz knock-out mice. Other receptors linked to Gi/o proteins [gamma-aminobutyric acid type B receptor (GABAB), adenosine A1 and muscarinic acetylcholine receptors] were not affected in Galphaz knock-out mice. These results suggest that the 5HT1A receptor may be linked to Galphaz protein, as reported previously in cell culture but shown here in an intact neural network.

Analysis of Knock Phenomenon Induced in a Constant Volume Chamber by Local Gas Temperature Measurement and Visualization

NASA Astrophysics Data System (ADS)

Moriyoshi, Yasuo; Kobayashi, Shigemi; Enomoto, Yoshiteru

Knock phenomenon in SI engines is regarded as an auto-ignition of unburned end-gas, and it has been widely examined by using rapid compression machines (RCM), shock-tubes or test engines. Recent researches point out the importance of the low temperature chemical reaction and the negative temperature coefficient (NTC). To investigate the effects, analyses of instantaneous local gas temperature, flow visualization and gaseous pressure were conducted in this study. As measurements using real engines are too difficult to analyze, the authors aimed to make measurements using a constant volume vessel under knock conditions where propagating flame exists during the induction time of auto-ignition. Adopting the two-wire thermocouple method enabled us to measure the instantaneous local gas temperature until the moment when the flame front passes by. High-speed images inside the unburned region were also recorded simultaneously using an endoscope. As a result, it was found that when knock occurs, the auto-ignition initiation time seems slightly early compared to the results without knock. This causes a higher volume ratio of unburned mixture and existence of many hot spots and stochastically leads to an initiation of knock.

78 FR 14091 - Public Availability of Equal Employment Opportunity Commission (EEOC) FY 2012 Service Contract...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-03-04

... EQUAL EMPLOYMENT OPPORTUNITY COMMISSION Public Availability of Equal Employment Opportunity Commission (EEOC) FY 2012 Service Contract Inventory AGENCY: Equal Employment Opportunity Commission. ACTION... Section 743 of Division C of the Consolidated Appropriations Act of 2010 (Pub. L. 111-117), Equal...

Efficient Knock-in of a Point Mutation in Porcine Fibroblasts Using the CRISPR/Cas9-GMNN Fusion Gene.

PubMed

Gerlach, Max; Kraft, Theresia; Brenner, Bernhard; Petersen, Björn; Niemann, Heiner; Montag, Judith

2018-06-13

During CRISPR/Cas9 mediated genome editing, site-specific double strand breaks are introduced and repaired either unspecific by non-homologous end joining (NHEJ) or sequence dependent by homology directed repair (HDR). Whereas NHEJ-based generation of gene knock-out is widely performed, the HDR-based knock-in of specific mutations remains a bottleneck. Especially in primary cell lines that are essential for the generation of cell culture and animal models of inherited human diseases, knock-in efficacy is insufficient and needs significant improvement. Here, we tested two different approaches to increase the knock-in frequency of a specific point mutation into the MYH7 -gene in porcine fetal fibroblasts. We added a small molecule inhibitor of NHEJ, SCR7 (5,6-bis((E)-benzylideneamino)-2-mercaptopyrimidin-4-ol), during genome editing and screened cell cultures for the point mutation. However, this approach did not yield increased knock-in rates. In an alternative approach, we fused humanized Cas9 (hCas9) to the N-terminal peptide of the Geminin gene ( GMNN ). The fusion protein is degraded in NHEJ-dominated cell cycle phases, which should increase HDR-rates. Using hCas9- GMNN and point mutation-specific real time PCR screening, we found a two-fold increase in genome edited cell cultures. This increase of HDR by hCas9- GMNN provides a promising way to enrich specific knock-in in porcine fibroblast cultures for somatic cloning approaches.

Knock-in of Enhanced Green Fluorescent Protein or/and Human Fibroblast Growth Factor 2 Gene into β-Casein Gene Locus in the Porcine Fibroblasts to Produce Therapeutic Protein.

PubMed

Lee, Sang Mi; Kim, Ji Woo; Jeong, Young-Hee; Kim, Se Eun; Kim, Yeong Ji; Moon, Seung Ju; Lee, Ji-Hye; Kim, Keun-Jung; Kim, Min-Kyu; Kang, Man-Jong

2014-11-01

Transgenic animals have become important tools for the production of therapeutic proteins in the domestic animal. Production efficiencies of transgenic animals by conventional methods as microinjection and retrovirus vector methods are low, and the foreign gene expression levels are also low because of their random integration in the host genome. In this study, we investigated the homologous recombination on the porcine β-casein gene locus using a knock-in vector for the β-casein gene locus. We developed the knock-in vector on the porcine β-casein gene locus and isolated knock-in fibroblast for nuclear transfer. The knock-in vector consisted of the neomycin resistance gene (neo) as a positive selectable marker gene, diphtheria toxin-A gene as negative selection marker, and 5' arm and 3' arm from the porcine β-casein gene. The secretion of enhanced green fluorescent protein (EGFP) was more easily detected in the cell culture media than it was by western blot analysis of cell extract of the HC11 mouse mammary epithelial cells transfected with EGFP knock-in vector. These results indicated that a knock-in system using β-casein gene induced high expression of transgene by the gene regulatory sequence of endogenous β-casein gene. These fibroblasts may be used to produce transgenic pigs for the production of therapeutic proteins via the mammary glands.

Knock-in of Enhanced Green Fluorescent Protein or/and Human Fibroblast Growth Factor 2 Gene into β-Casein Gene Locus in the Porcine Fibroblasts to Produce Therapeutic Protein

PubMed Central

Lee, Sang Mi; Kim, Ji Woo; Jeong, Young-Hee; Kim, Se Eun; Kim, Yeong Ji; Moon, Seung Ju; Lee, Ji-Hye; Kim, Keun-Jung; Kim, Min-Kyu; Kang, Man-Jong

2014-01-01

Transgenic animals have become important tools for the production of therapeutic proteins in the domestic animal. Production efficiencies of transgenic animals by conventional methods as microinjection and retrovirus vector methods are low, and the foreign gene expression levels are also low because of their random integration in the host genome. In this study, we investigated the homologous recombination on the porcine β-casein gene locus using a knock-in vector for the β-casein gene locus. We developed the knock-in vector on the porcine β-casein gene locus and isolated knock-in fibroblast for nuclear transfer. The knock-in vector consisted of the neomycin resistance gene (neo) as a positive selectable marker gene, diphtheria toxin-A gene as negative selection marker, and 5′ arm and 3′ arm from the porcine β-casein gene. The secretion of enhanced green fluorescent protein (EGFP) was more easily detected in the cell culture media than it was by western blot analysis of cell extract of the HC11 mouse mammary epithelial cells transfected with EGFP knock-in vector. These results indicated that a knock-in system using β-casein gene induced high expression of transgene by the gene regulatory sequence of endogenous β-casein gene. These fibroblasts may be used to produce transgenic pigs for the production of therapeutic proteins via the mammary glands. PMID:25358326

Targeted Knock-Down of miR21 Primary Transcripts Using snoMEN Vectors Induces Apoptosis in Human Cancer Cell Lines.

PubMed

Ono, Motoharu; Yamada, Kayo; Avolio, Fabio; Afzal, Vackar; Bensaddek, Dalila; Lamond, Angus I

2015-01-01

We have previously reported an antisense technology, 'snoMEN vectors', for targeted knock-down of protein coding mRNAs using human snoRNAs manipulated to contain short regions of sequence complementarity with the mRNA target. Here we characterise the use of snoMEN vectors to target the knock-down of micro RNA primary transcripts. We document the specific knock-down of miR21 in HeLa cells using plasmid vectors expressing miR21-targeted snoMEN RNAs and show this induces apoptosis. Knock-down is dependent on the presence of complementary sequences in the snoMEN vector and the induction of apoptosis can be suppressed by over-expression of miR21. Furthermore, we have also developed lentiviral vectors for delivery of snoMEN RNAs and show this increases the efficiency of vector transduction in many human cell lines that are difficult to transfect with plasmid vectors. Transduction of lentiviral vectors expressing snoMEN targeted to pri-miR21 induces apoptosis in human lung adenocarcinoma cells, which express high levels of miR21, but not in human primary cells. We show that snoMEN-mediated suppression of miRNA expression is prevented by siRNA knock-down of Ago2, but not by knock-down of Ago1 or Upf1. snoMEN RNAs colocalise with Ago2 in cell nuclei and nucleoli and can be co-immunoprecipitated from nuclear extracts by antibodies specific for Ago2.

Public Notice: 2016-03, In the matter of Redwood ERC

EPA Pesticide Factsheets

Under the provisions of the Clean Water Act, Resource Conservation And Recovery Act, Safe Drinking Water Act and Toxic Substances Control Act, EPA is providing the opportunity for public comment on the proposed actions listed below.

Oversight Hearing on the Equal Employment Opportunity Commission's Implementation of the Americans with Disabilities Act (Title I on Employment and Title V Covering Miscellaneous Provisions). Hearing before the Subcommittee on Employment Opportunities of the Committee on Education and Labor, House of Representatives, One Hundred Second Congress, First Session (October 30, 1991).

ERIC Educational Resources Information Center

Congress of the U.S., Washington, DC. House Committee on Education and Labor.

This hearing transcript presents testimony, a prepared statement, and supplemental materials provided by Evan J. Kemp, Jr., Chairman of the Equal Employment Opportunities Commission (EEOC), specifically related to implementation of the Americans with Disabilities Act (ADA). The hearing deals with EEOC enforcement strategies for implementing Title…

CRISPR/Cas9 Allows Efficient and Complete Knock-In of a Destabilization Domain-Tagged Essential Protein in a Human Cell Line, Allowing Rapid Knockdown of Protein Function

PubMed Central

Park, Arnold; Won, Sohui T.; Pentecost, Mickey; Bartkowski, Wojciech; Lee, Benhur

2014-01-01

Although modulation of protein levels is an important tool for study of protein function, it is difficult or impossible to knockdown or knockout genes that are critical for cell growth or viability. For such genes, a conditional knockdown approach would be valuable. The FKBP protein-based destabilization domain (DD)-tagging approach, which confers instability to the tagged protein in the absence of the compound Shield-1, has been shown to provide rapid control of protein levels determined by Shield-1 concentration. Although a strategy to knock-in DD-tagged protein at the endogenous loci has been employed in certain parasite studies, partly due to the relative ease of knock-in as a result of their mostly haploid lifecycles, this strategy has not been demonstrated in diploid or hyperploid mammalian cells due to the relative difficulty of achieving complete knock-in in all alleles. The recent advent of CRISPR/Cas9 homing endonuclease-mediated targeted genome cleavage has been shown to allow highly efficient homologous recombination at the targeted locus. We therefore assessed the feasibility of using CRISPR/Cas9 to achieve complete knock-in to DD-tag the essential gene Treacher Collins-Franceschetti syndrome 1 (TCOF1) in human 293T cells. Using a double antibiotic selection strategy to select clones with at least two knock-in alleles, we obtained numerous complete knock-in clones within three weeks of initial transfection. DD-TCOF1 expression in the knock-in cells was Shield-1 concentration-dependent, and removal of Shield-1 resulted in destabilization of DD-TCOF1 over the course of hours. We further confirmed that the tagged TCOF1 retained the nucleolar localization of the wild-type untagged protein, and that destabilization of DD-TCOF1 resulted in impaired cell growth, as expected for a gene implicated in ribosome biogenesis. CRISPR/Cas9-mediated homologous recombination to completely knock-in a DD tag likely represents a generalizable and efficient strategy to achieve rapid modulation of protein levels in mammalian cells. PMID:24743236

CRISPR/Cas9 allows efficient and complete knock-in of a destabilization domain-tagged essential protein in a human cell line, allowing rapid knockdown of protein function.

PubMed

Park, Arnold; Won, Sohui T; Pentecost, Mickey; Bartkowski, Wojciech; Lee, Benhur

2014-01-01

Although modulation of protein levels is an important tool for study of protein function, it is difficult or impossible to knockdown or knockout genes that are critical for cell growth or viability. For such genes, a conditional knockdown approach would be valuable. The FKBP protein-based destabilization domain (DD)-tagging approach, which confers instability to the tagged protein in the absence of the compound Shield-1, has been shown to provide rapid control of protein levels determined by Shield-1 concentration. Although a strategy to knock-in DD-tagged protein at the endogenous loci has been employed in certain parasite studies, partly due to the relative ease of knock-in as a result of their mostly haploid lifecycles, this strategy has not been demonstrated in diploid or hyperploid mammalian cells due to the relative difficulty of achieving complete knock-in in all alleles. The recent advent of CRISPR/Cas9 homing endonuclease-mediated targeted genome cleavage has been shown to allow highly efficient homologous recombination at the targeted locus. We therefore assessed the feasibility of using CRISPR/Cas9 to achieve complete knock-in to DD-tag the essential gene Treacher Collins-Franceschetti syndrome 1 (TCOF1) in human 293T cells. Using a double antibiotic selection strategy to select clones with at least two knock-in alleles, we obtained numerous complete knock-in clones within three weeks of initial transfection. DD-TCOF1 expression in the knock-in cells was Shield-1 concentration-dependent, and removal of Shield-1 resulted in destabilization of DD-TCOF1 over the course of hours. We further confirmed that the tagged TCOF1 retained the nucleolar localization of the wild-type untagged protein, and that destabilization of DD-TCOF1 resulted in impaired cell growth, as expected for a gene implicated in ribosome biogenesis. CRISPR/Cas9-mediated homologous recombination to completely knock-in a DD tag likely represents a generalizable and efficient strategy to achieve rapid modulation of protein levels in mammalian cells.

A knock-in/knock-out mouse model of HSPB8-associated distal hereditary motor neuropathy and myopathy reveals toxic gain-of-function of mutant Hspb8.

PubMed

Bouhy, Delphine; Juneja, Manisha; Katona, Istvan; Holmgren, Anne; Asselbergh, Bob; De Winter, Vicky; Hochepied, Tino; Goossens, Steven; Haigh, Jody J; Libert, Claude; Ceuterick-de Groote, Chantal; Irobi, Joy; Weis, Joachim; Timmerman, Vincent

2018-01-01

Mutations in the small heat shock protein B8 gene (HSPB8/HSP22) have been associated with distal hereditary motor neuropathy, Charcot-Marie-Tooth disease, and recently distal myopathy. It is so far not clear how mutant HSPB8 induces the neuronal and muscular phenotypes and if a common pathogenesis lies behind these diseases. Growing evidence points towards a role of HSPB8 in chaperone-associated autophagy, which has been shown to be a determinant for the clearance of poly-glutamine aggregates in neurodegenerative diseases but also for the maintenance of skeletal muscle myofibrils. To test this hypothesis and better dissect the pathomechanism of mutant HSPB8, we generated a new transgenic mouse model leading to the expression of the mutant protein (knock-in lines) or the loss-of-function (functional knock-out lines) of the endogenous protein Hspb8. While the homozygous knock-in mice developed motor deficits associated with degeneration of peripheral nerves and severe muscle atrophy corroborating patient data, homozygous knock-out mice had locomotor performances equivalent to those of wild-type animals. The distal skeletal muscles of the post-symptomatic homozygous knock-in displayed Z-disk disorganisation, granulofilamentous material accumulation along with Hspb8, αB-crystallin (HSPB5/CRYAB), and desmin aggregates. The presence of the aggregates correlated with reduced markers of effective autophagy. The sciatic nerve of the homozygous knock-in mice was characterized by low autophagy potential in pre-symptomatic and Hspb8 aggregates in post-symptomatic animals. On the other hand, the sciatic nerve of the homozygous knock-out mice presented a normal morphology and their distal muscle displayed accumulation of abnormal mitochondria but intact myofiber and Z-line organisation. Our data, therefore, suggest that toxic gain-of-function of mutant Hspb8 aggregates is a major contributor to the peripheral neuropathy and the myopathy. In addition, mutant Hspb8 induces impairments in autophagy that may aggravate the phenotype.

75 FR 31787 - Agency Information Collection Activities: Existing Collection; Emergency Extension

Federal Register 2010, 2011, 2012, 2013, 2014

2010-06-04

... EQUAL EMPLOYMENT OPPORTUNITY COMMISSION Agency Information Collection Activities: Existing Collection; Emergency Extension AGENCY: Equal Employment Opportunity Commission. ACTION: Notice of... Paperwork Reduction Act, the Equal Employment Opportunity Commission (EEOC or Commission) announces that it...

7 CFR 1951.231 - Special provisions applicable to Economic Opportunity (EO) Cooperative Loans.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 7 Agriculture 14 2010-01-01 2009-01-01 true Special provisions applicable to Economic Opportunity... Special provisions applicable to Economic Opportunity (EO) Cooperative Loans. (a) Withdrawal of member and... for Economic Opportunity Cooperative loans must be handled under the Federal Claims Collection Act...

20 CFR 664.610 - How is the summer employment opportunities element administered?

Code of Federal Regulations, 2012 CFR

2012-04-01

... 20 Employees' Benefits 4 2012-04-01 2012-04-01 false How is the summer employment opportunities..., DEPARTMENT OF LABOR (CONTINUED) YOUTH ACTIVITIES UNDER TITLE I OF THE WORKFORCE INVESTMENT ACT Summer Employment Opportunities § 664.610 How is the summer employment opportunities element administered? Chief...

20 CFR 664.610 - How is the summer employment opportunities element administered?

Code of Federal Regulations, 2013 CFR

2013-04-01

... 20 Employees' Benefits 4 2013-04-01 2013-04-01 false How is the summer employment opportunities..., DEPARTMENT OF LABOR (CONTINUED) YOUTH ACTIVITIES UNDER TITLE I OF THE WORKFORCE INVESTMENT ACT Summer Employment Opportunities § 664.610 How is the summer employment opportunities element administered? Chief...

20 CFR 664.610 - How is the summer employment opportunities element administered?

Code of Federal Regulations, 2014 CFR

2014-04-01

... 20 Employees' Benefits 4 2014-04-01 2014-04-01 false How is the summer employment opportunities..., DEPARTMENT OF LABOR (CONTINUED) YOUTH ACTIVITIES UNDER TITLE I OF THE WORKFORCE INVESTMENT ACT Summer Employment Opportunities § 664.610 How is the summer employment opportunities element administered? Chief...

20 CFR 664.610 - How is the summer employment opportunities element administered?

Code of Federal Regulations, 2011 CFR

2011-04-01

... 20 Employees' Benefits 3 2011-04-01 2011-04-01 false How is the summer employment opportunities..., DEPARTMENT OF LABOR YOUTH ACTIVITIES UNDER TITLE I OF THE WORKFORCE INVESTMENT ACT Summer Employment Opportunities § 664.610 How is the summer employment opportunities element administered? Chief elected officials...

20 CFR 664.610 - How is the summer employment opportunities element administered?

Code of Federal Regulations, 2010 CFR

2010-04-01

... 20 Employees' Benefits 3 2010-04-01 2010-04-01 false How is the summer employment opportunities..., DEPARTMENT OF LABOR YOUTH ACTIVITIES UNDER TITLE I OF THE WORKFORCE INVESTMENT ACT Summer Employment Opportunities § 664.610 How is the summer employment opportunities element administered? Chief elected officials...

Estrogen-Related Receptor Alpha Modulates Lactate Dehydrogenase Activity in Thyroid Tumors

PubMed Central

Mirebeau-Prunier, Delphine; Le Pennec, Soazig; Jacques, Caroline; Fontaine, Jean-Fred; Gueguen, Naig; Boutet-Bouzamondo, Nathalie; Donnart, Audrey; Malthièry, Yves; Savagner, Frédérique

2013-01-01

Metabolic modifications of tumor cells are hallmarks of cancer. They exhibit an altered metabolism that allows them to sustain higher proliferation rates in hostile environment outside the cell. In thyroid tumors, the expression of the estrogen-related receptor α (ERRα), a major factor of metabolic adaptation, is closely related to the oxidative metabolism and the proliferative status of the cells. To elucidate the role played by ERRα in the glycolytic adaptation of tumor cells, we focused on the regulation of lactate dehydrogenases A and B (LDHA, LDHB) and the LDHA/LDHB ratio. Our study included tissue samples from 10 classical and 10 oncocytic variants of follicular thyroid tumors and 10 normal thyroid tissues, as well as samples from three human thyroid tumor cell lines: FTC-133, XTC.UC1 and RO82W-1. We identified multiple cis-acting promoter elements for ERRα, in both the LDHA and LDHB genes. The interaction between ERRα and LDH promoters was confirmed by chromatin immunoprecipitation assays and in vitro analysis for LDHB. Using knock-in and knock-out cellular models, we found an inverse correlation between ERRα expression and LDH activity. This suggests that thyroid tumor cells may reprogram their metabolic pathways through the up-regulation of ERRα by a process distinct from that proposed by the recently revisited Warburg hypothesis. PMID:23516535

Mutations in the murine homologue of TUBB5 cause microcephaly by perturbing cell cycle progression and inducing p53-associated apoptosis.

PubMed

Breuss, Martin; Fritz, Tanja; Gstrein, Thomas; Chan, Kelvin; Ushakova, Lyubov; Yu, Nuo; Vonberg, Frederick W; Werner, Barbara; Elling, Ulrich; Keays, David A

2016-04-01

Microtubules play a crucial role in the generation, migration and differentiation of nascent neurons in the developing vertebrate brain. Mutations in the constituents of microtubules, the tubulins, are known to cause an array of neurological disorders, including lissencephaly, polymicrogyria and microcephaly. In this study we explore the genetic and cellular mechanisms that cause TUBB5-associated microcephaly by exploiting two new mouse models: a conditional E401K knock-in, and a conditional knockout animal. These mice present with profound microcephaly due to a loss of upper-layer neurons that correlates with massive apoptosis and upregulation of p53. This phenotype is associated with a delay in cell cycle progression and ectopic DNA elements in progenitors, which is dependent on the dosage of functional Tubb5. Strikingly, we report ectopic Sox2-positive progenitors and defects in spindle orientation in our knock-in mouse line, which are absent in knockout animals. This work sheds light on the functional repertoire of Tubb5, reveals that the E401K mutation acts by a complex mechanism, and demonstrates that the cellular pathology driving TUBB5-associated microcephaly is cell death. © 2016. Published by The Company of Biologists Ltd.

Subunits of the Drosophila Actin-Capping Protein Heterodimer Regulate Each Other at Multiple Levels

PubMed Central

Amândio, Ana Rita; Gaspar, Pedro; Whited, Jessica L.; Janody, Florence

2014-01-01

The actin-Capping Protein heterodimer, composed of the α and β subunits, is a master F-actin regulator. In addition to its role in many cellular processes, Capping Protein acts as a main tumor suppressor module in Drosophila and in humans, in part, by restricting the activity of Yorkie/YAP/TAZ oncogenes. We aimed in this report to understand how both subunits regulate each other in vivo. We show that the levels and capping activities of both subunits must be tightly regulated to control F-actin levels and consequently growth of the Drosophila wing. Overexpressing capping protein α and β decreases both F-actin levels and tissue growth, while expressing forms of Capping Protein that have dominant negative effects on F-actin promote tissue growth. Both subunits regulate each other's protein levels. In addition, overexpressing one of the subunit in tissues knocked-down for the other increases the mRNA and protein levels of the subunit knocked-down and compensates for its loss. We propose that the ability of the α and β subunits to control each other's levels assures that a pool of functional heterodimer is produced in sufficient quantities to restrict the development of tumor but not in excess to sustain normal tissue growth. PMID:24788460

Workforce Investment Act Youth Provisions and School-to-Work Opportunities: A Context for Collaboration.

ERIC Educational Resources Information Center

Spangler, D.

This document is intended to help existing school-to-work (STW) partnerships and the new local work force investment boards understand the opportunities for achieving common ground to better serve youth during implementation of the Workforce Investment Act (WIA) and formation of the new youth councils. The following are among the topics discussed:…

Workforce Innovation and Opportunity Act: Information on Planned Changes to State Performance Reporting and Related Challenges. Report to Congressional Committees. GAO-16-287

ERIC Educational Resources Information Center

US Government Accountability Office, 2016

2016-01-01

Enacted in 2014, the Workforce Innovation and Opportunity Act (WIOA) brought numerous changes to existing federal employment and training programs, including requiring the Department of Labor (DOL) and the Department of Education (Education) to implement a common performance accountability system across the six WIOA-designated core programs. WIOA…

The Personal Responsibility and Work Opportunity Act of 1996: What Welfare Reform Means for Head Start.

ERIC Educational Resources Information Center

Shuell, Julie; Hanna, Jeff; Oterlei, Jannell; Kariger, Patricia

This National Head Start Association booklet outlines the main provisions of the Personal Responsibility and Work Opportunity Act and describes how it may affect local Head Start Programs. The document is intended to serve as a starting point for local programs, parents, administrators and policy workers to discuss and plan how Head Start will…

Equal Educational Opportunity: The State of the Law. ERIC-CUE Urban Disadvantaged Series, Number 48.

ERIC Educational Resources Information Center

Glickstein, Howard A.

This paper addresses the state of the law of equal educational opportunity. Among the laws, acts, and statutes addressed are the following: the Fourteenth Amendment to the U.S. Constitution, the implementation of school desegregation in the North and South, the Civil Rights Act of 1964, Title IX of the Education Amendments of 1972, the Educational…

77 FR 23791 - Notice With Respect to List of Countries Denying Fair Market Opportunities for Government-Funded...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-04-20

... Airway Improvement Act of 1982, as amended (49 U.S.C. 50104), the United States Trade Representative (USTR) has determined not to list any countries as denying fair market opportunities for U.S. products...: Section 533 of the Airport and Airway Improvement Act of 1982, as amended by section 115 of the Airport...

76 FR 24082 - Notice With Respect to List of Countries Denying Fair Market Opportunities for Government-Funded...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-04-29

... Airway Improvement Act of 1982, as amended (49 U.S.C. 50104), the United States Trade Representative (USTR) has determined not to list any countries as denying fair market opportunities for U.S. products...: Section 533 of the Airport and Airway Improvement Act of 1982, as amended by section 115 of the Airport...

78 FR 25337 - Notice With Respect to List of Countries Denying Fair Market Opportunities for Government-Funded...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-04-30

... Airway Improvement Act of 1982, as amended (49 U.S.C. 50104), the United States Trade Representative (USTR) has determined not to list any countries as denying fair market opportunities for U.S. products... INFORMATION: Section 533 of the Airport and Airway Improvement Act of 1982, as amended by section 115 of the...

The effect of SIRT1 protein knock down on PGC-1α acetylation during skeletal muscle contraction.

PubMed

Park, Dae Ryoung; Kim, Jeong Seok; Kim, Chang Keun

2014-03-01

The purpose of this study was to investigate the effect of Sirtuin 1 (SIRT1) and General control nonderepressible 5 (GCN5) knock down on peroxisome proliferator- activated receptor gamma coactivator 1-alpha (PGC-1α) deacetylation during electrical stimulated skeletal muscle contraction. Skeletal muscle primary cell were isolated from C57BL/6 mice gastrocnemius and transfected lentiviral SIRT1 and GCN5 shRNA. Knock downed muscle cell were stimulated by electrical stimulation (1Hz, 3min) and collected for PGC-1α deceatylation assays. Immunoprecipitation performed for PGC-1α deacetylation, acetyl-lysine level was measured. Our resulted showed SIRT1 knock down not influenced to PGC-1α deacetylation during electrical stimulation induced muscle contraction while GCN5 knock down decreased PGC-1α deacetylation significantly (p<0.05). This study can be concluded that GCN5 is a critical factor for muscle contraction induced PGC-1α deacetylation.

The Full Employment and Balanced Growth Act of 1977; Authorization of Appropriations for CETA, and the President's Economic Stimulus Proposals; Hearings before the Subcommittee on Employment Opportunities of the Committee on Education and Labor, House of Representatives, Ninety-Fifth Congress, First Session on H.R. 50 and H.R. 2992, Volume 1.

ERIC Educational Resources Information Center

Congress of the U.S., Washington, DC. House Committee on Education and Labor.

The proceedings from the 2 days of hearings before the Subcommittee on Employment Opportunities on the Full Employment and Balanced Growth Act of 1977, the authorizations and appropriations for the Comprehensive Employment and Training Act (CETA), and the President's economic stimulus proposals are presented in this report. All prepared…

The Reduced Effectiveness of EGR to Mitigate Knock at High Loads in Boosted SI Engines

DOE Office of Scientific and Technical Information (OSTI.GOV)

Szybist, James P.; Wagnon, Scott W.; Splitter, Derek A.

Numerous studies have demonstrated that exhaust gas recirculation (EGR) can attenuate knock propensity in spark ignition (SI) engines at naturally aspirated or lightly boosted conditions. In this paper, we investigate the role of cooled EGR under higher load conditions with multiple fuel compositions, where highly retarded combustion phasing typical of modern SI engines was used. It was found that under these conditions, EGR attenuation of knock is greatly reduced, where EGR doesn’t allow significant combustion phasing advance as it does under lighter load conditions. Detailed combustion analysis shows that when EGR is added, the polytropic coefficient increases causing the compressivemore » pressure and temperature to increase. At sufficiently highly boosted conditions, the increase in polytropic coefficient and additional trapped mass from EGR can sufficiently reduce fuel ignition delay to overcome knock attenuation effects. Kinetic modeling demonstrates that the effectiveness of EGR to mitigate knock is highly dependent on the pressure-temperature condition. Experiments at 2000 rpm have confirmed reduced fuel ignition delay under highly boosted conditions relevant to modern downsized boosted SI engines, where in-cylinder pressure is higher and the temperature is cooler. Finally, at these conditions, charge reactivity increases compared to naturally aspirated conditions, and attenuation of knock by EGR is reduced.« less

The Reduced Effectiveness of EGR to Mitigate Knock at High Loads in Boosted SI Engines

DOE PAGES

Szybist, James P.; Wagnon, Scott W.; Splitter, Derek A.; ...

2017-09-04

Numerous studies have demonstrated that exhaust gas recirculation (EGR) can attenuate knock propensity in spark ignition (SI) engines at naturally aspirated or lightly boosted conditions. In this paper, we investigate the role of cooled EGR under higher load conditions with multiple fuel compositions, where highly retarded combustion phasing typical of modern SI engines was used. It was found that under these conditions, EGR attenuation of knock is greatly reduced, where EGR doesn’t allow significant combustion phasing advance as it does under lighter load conditions. Detailed combustion analysis shows that when EGR is added, the polytropic coefficient increases causing the compressivemore » pressure and temperature to increase. At sufficiently highly boosted conditions, the increase in polytropic coefficient and additional trapped mass from EGR can sufficiently reduce fuel ignition delay to overcome knock attenuation effects. Kinetic modeling demonstrates that the effectiveness of EGR to mitigate knock is highly dependent on the pressure-temperature condition. Experiments at 2000 rpm have confirmed reduced fuel ignition delay under highly boosted conditions relevant to modern downsized boosted SI engines, where in-cylinder pressure is higher and the temperature is cooler. Finally, at these conditions, charge reactivity increases compared to naturally aspirated conditions, and attenuation of knock by EGR is reduced.« less

Paired immunoglobulin-like receptor B knockout does not enhance axonal regeneration or locomotor recovery after spinal cord injury.

PubMed

Nakamura, Yuka; Fujita, Yuki; Ueno, Masaki; Takai, Toshiyuki; Yamashita, Toshihide

2011-01-21

Myelin components that inhibit axonal regeneration are believed to contribute significantly to the lack of axonal regeneration noted in the adult central nervous system. Three proteins found in myelin, Nogo, myelin-associated glycoprotein, and oligodendrocyte-myelin glycoprotein, inhibit neurite outgrowth in vitro. All of these proteins interact with the same receptors, namely, the Nogo receptor (NgR) and paired immunoglobulin-like receptor B (PIR-B). As per previous reports, corticospinal tract (CST) regeneration is not enhanced in NgR-knock-out mice after spinal cord injury. Therefore, we assessed CST regeneration in PIR-B-knock-out mice. We found that hindlimb motor function, as assessed using the Basso mouse scale, footprint test, inclined plane test, and beam walking test, did not differ between the PIR-B-knock-out and wild-type mice after dorsal hemisection of the spinal cord. Further, tracing of the CST fibers after injury did not reveal enhanced axonal regeneration or sprouting in the CST of the PIR-B-knock-out mice. Systemic administration of NEP1-40, a NgR antagonist, to PIR-B knock-out mice did not enhance the regenerative response. These results indicate that PIR-B knock-out is not sufficient to induce extensive axonal regeneration after spinal cord injury.

Analysis of Spark-Ignition Engine Knock as Seen in Photographs Taken at 200,000 Frames Per Second

NASA Technical Reports Server (NTRS)

Miller, Cearcy D.; Olsen, H. Lowell; Logan, Walter O., Jr.; Osterstrom, Gordon E

1946-01-01

A motion picture of the development of knock in a spark-ignition engine, is presented, which consists of 20 photographs taken at intervals of 5 microseconds, or at a rate of 200,000 photographs a second, with an equivalent wide-open exposure time of 6.4 microseconds for each photograph. A motion picture of a complete combustion process, including the development of knock, taken at the rate of 40,000 photographs a second is also presented to assist the reader in orienting the photographs of the knock development taken at 200,000 frames per second. The photographs taken at 200,000 frames per second are analyzed and the conclusion is made that the type of knock in the spark-ignition engine involving violent gas vibration originates as self-propagating disturbance starting at a point in the.burn1ig or autoigniting gases and spreading out from that point through the incompletely burned gases at a rate as high as 6800 feet per second, or about twice the speed of sound in the burned gases. Apparent formation of free carbon particles in both the burning and the burned gas is observed within 10 microseconds after passage of the knock disturbance through the gases.

Novel In Vivo Model for Combinatorial Fluorescence Labeling in Mouse Prostate

PubMed Central

Fang, Xiaolan; Gyabaah, Kenneth; Nickkholgh, Bita; Cline, J. Mark; Balaji, K.C.

2015-01-01

BACKGROUND The epithelial layer of prostate glands contains several types of cells, including luminal and basal cells. Yet there is paucity of animal models to study the cellular origin of normal or neoplastic development in the prostate to facilitate the treatment of heterogenous prostate diseases by targeting individual cell lineages. METHODS We developed a mouse model that expresses different types of fluorescent proteins (XFPs) specifically in prostatic cells. Using an in vivo stochastic fluorescent protein combinatorial strategy, XFP signals were expressed specifically in prostate of Protein Kinase D1 (PKD1) knock-out, K-RasG12D knock-in, and Phosphatase and tensin homolog (PTEN) and PKD1 double knock-out mice under the control of PB-Cre promoter. RESULTS In vivo XFP signals were observed in prostate of PKD1 knock-out, K-RasG12D knock-in, and PTEN PKD1 double knock-out mice, which developed normal, hyperplastic, and neoplastic prostate, respectively. The patchy expression pattern of XFPs in neoplasia tissue indicated the clonal origin of cancer cells in the prostate. CONCLUSIONS The transgenic mouse models demonstrate combinatorial fluorescent protein expression in normal and cancerous prostatic tissues. This novel prostate-specific fluorescent labeled mouse model, which we named Prorainbow, could be useful in studying benign and malignant pathology of prostate. PMID:25753731

Novel In Vivo model for combinatorial fluorescence labeling in mouse prostate.

PubMed

Fang, Xiaolan; Gyabaah, Kenneth; Nickkholgh, Bita; Cline, J Mark; Balaji, K C

2015-06-15

The epithelial layer of prostate glands contains several types of cells, including luminal and basal cells. Yet there is paucity of animal models to study the cellular origin of normal or neoplastic development in the prostate to facilitate the treatment of heterogenous prostate diseases by targeting individual cell lineages. We developed a mouse model that expresses different types of fluorescent proteins (XFPs) specifically in prostatic cells. Using an in vivo stochastic fluorescent protein combinatorial strategy, XFP signals were expressed specifically in prostate of Protein Kinase D1 (PKD1) knock-out, K-Ras(G) (12) (D) knock-in, and Phosphatase and tensin homolog (PTEN) and PKD1 double knock-out mice under the control of PB-Cre promoter. In vivo XFP signals were observed in prostate of PKD1 knock-out, K-Ras(G) (12) (D) knock-in, and PTEN PKD1 double knock-out mice, which developed normal, hyperplastic, and neoplastic prostate, respectively. The patchy expression pattern of XFPs in neoplasia tissue indicated the clonal origin of cancer cells in the prostate. The transgenic mouse models demonstrate combinatorial fluorescent protein expression in normal and cancerous prostatic tissues. This novel prostate-specific fluorescent labeled mouse model, which we named Prorainbow, could be useful in studying benign and malignant pathology of prostate. © 2015 Wiley Periodicals, Inc.

Exploring the Relationship Between Octane Sensitivity and Heat-of-Vaporization

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sluder, C. Scott; Szybist, James P.; McCormick, Robert L.

2016-04-05

The latent heat-of-vaporization (HoV) of blends of biofuel and hydrocarbon components into gasolines has recently experienced expanded interest because of the potential for increased HoV to increase fuel knock resistance in direct-injection (DI) engines. Several studies have been conducted, with some studies identifying an additional anti-knock benefit from HoV and others failing to arrive at the same conclusion. Consideration of these studies holistically shows that they can be grouped according to the level of fuel octane sensitivity variation within their fuel matrices. When comparing fuels of different octane sensitivity significant additional anti-knock benefits associated with HoV are sometimes observed. Studiesmore » that fix the octane sensitivity find that HoV does not produce additional anti-knock benefit. New studies were performed at ORNL and NREL to further investigate the relationship between HoV and octane sensitivity. Three fuels were formulated for the ORNL study with matched RON and octane sensitivity, but with differing HoV. Experiments with these fuels in a 1.6-liter GTDI engine showed that the fuels exhibited very similar combustion phasing under knock-limited spark advance (KLSA) conditions. Fuels having a range of RON, octane sensitivity, and HoV were tested at NREL in a single-cylinder GDI engine under conditions where octane sensitivity has little effect on knock resistance. KLSA was found to be well correlated with RON. These results reinforce the concept that HoV anti-knock effects can be viewed as a contributor to octane sensitivity. From this viewpoint, HoV effects manifest themselves as increases in octane sensitivity.« less

The knock-down of the expression of MdMLO19 reduces susceptibility to powdery mildew (Podosphaera leucotricha) in apple (Malus domestica).

PubMed

Pessina, Stefano; Angeli, Dario; Martens, Stefan; Visser, Richard G F; Bai, Yuling; Salamini, Francesco; Velasco, Riccardo; Schouten, Henk J; Malnoy, Mickael

2016-10-01

Varieties resistant to powdery mildew (PM; caused by Podosphaera leucotricha) are a major component of sustainable apple production. Resistance can be achieved by knocking-out susceptibility S-genes to be singled out among members of the MLO (Mildew Locus O) gene family. Candidates are MLO S-genes of phylogenetic clade V up-regulated upon PM inoculation, such as MdMLO11 and 19 (clade V) and MdMLO18 (clade VII). We report the knock-down through RNA interference of MdMLO11 and 19, as well as the complementation of resistance with MdMLO18 in the Arabidopsis thaliana triple mlo mutant Atmlo2/6/12. The knock-down of MdMLO19 reduced PM disease severity by 75%, whereas the knock-down of MdMLO11, alone or in combination with MdMLO19, did not result in any reduction or additional reduction of susceptibility compared with MdMLO19 alone. The test in A. thaliana excluded a role for MdMLO18 in PM susceptibility. Cell wall appositions (papillae) were present in both PM-resistant and PM-susceptible plants, but were larger in resistant lines. No obvious negative phenotype was observed in plants with mlo genes knocked down. Apparently, MdMLO19 plays the pivotal role in apple PM susceptibility and its knock-down induces a very significant level of resistance. © 2016 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

Knock knee and the gait of six-year-old children.

PubMed

Pretkiewicz-Abacjew, E

2003-06-01

Knock knee (genu valgum) interferes with the locomotive and supporting function of the lower limb. In static conditions the load-bearing axis of the valgus limb is displaced laterally in relation to the middle of the joint, causing the knee joint, the ankle joint, and the foot as a whole to be weighted in the wrong way. The purpose of this work is to examine the influence of knock knee on gait kinematics. The gait of twenty-two 6-year-old children of both sexes in whom knock knee had been medically diagnosed was compared with the gait of 33 children of the same age whose knee joints conformed to the norm in formation and position. Gait was recorded separately for the sagittal and the frontal planes, using a video-computer system. The results of the examination indicated statistically significant differences in the gait of the two groups of children. These differences related mainly to the time features of gait and to data on the angles in the knee and ankle joints. Although the results obtained for other features of gait did not reveal statistical differences, these did indicate that the children with knock knee walked more slowly and with a lower cadence. The results indicate that knock knee in 6-year-old children has an adverse impact on the mechanics of the lower limb joints in gait and causes a deterioration in gait quality. Thus knock knee in children should not be treated merely as a superficial defect but should be subject to therapy and, more importantly, taken into account when introducing children to early sports training.

28 CFR 42.700 - Purpose.

Code of Federal Regulations, 2014 CFR

2014-07-01

... Administration DEPARTMENT OF JUSTICE NONDISCRIMINATION; EQUAL EMPLOYMENT OPPORTUNITY; POLICIES AND PROCEDURES... Discrimination Act of 1975 General Provisions § 42.700 Purpose. (a) This subpart implements the Age Discrimination Act of 1975, as amended (42 U.S.C. 5101-6107) (Act). Subject to certain exceptions, the Act...

Characterization of the first knock-out aldh7a1 zebrafish model for pyridoxine-dependent epilepsy using CRISPR-Cas9 technology

PubMed Central

Zabinyakov, Nikita; Bullivant, Garrett; Cao, Feng; Fernandez Ojeda, Matilde; Jia, Zheng Ping; Wen, Xiao-Yan; Dowling, James J.; Salomons, Gajja S.

2017-01-01

Pyridoxine dependent epilepsy (PDE) is caused by likely pathogenic variants in ALDH7A1 (PDE-ALDH7A1) and inherited autosomal recessively. Neurotoxic alpha-amino adipic semialdehyde (alpha-AASA), piperideine 6-carboxylate and pipecolic acid accumulate in body fluids. Neonatal or infantile onset seizures refractory to anti-epileptic medications are clinical features. Treatment with pyridoxine, arginine and lysine-restricted diet does not normalize neurodevelopmental outcome or accumulation of neurotoxic metabolites. There is no animal model for high throughput drug screening. For this reason, we developed and characterized the first knock-out aldh7a1 zebrafish model using CRISPR-Cas9 technology. Zebrafish aldh7a1 mutants were generated by using a vector free method of CRISPR-Cas9 mutagenesis. Genotype analysis of aldh7a1 knock-out zebrafish was performed by high resolution melt analysis, direct sequencing and QIAxcel system. Electroencephalogram was performed. Alpha-AASA, piperideine 6-carboxylate and pipecolic acid, were measured by liquid chromatography-tandem mass spectrometry. Our knock-out aldh7a1 zebrafish has homozygous 5 base pair (bp) mutation in ALDH7A1. Knock-out aldh7a1 embryos have spontaneous rapid increase in locomotion and a rapid circling swim behavior earliest 8-day post fertilization (dpf). Electroencephalogram revealed large amplitude spike discharges compared to wild type. Knock-out aldh7a1 embryos have elevated alpha-AASA, piperideine 6-carboxylate and pipecolic acid compared to wild type embryos at 3 dpf. Knock-out aldh7a1 embryos showed no aldh7a1 protein by western blot compared to wild type. Our knock-out aldh7a1 zebrafish is a well characterized model for large-scale drug screening using behavioral and biochemical features and accurately recapitulates the human PDE-ALDH7A1 disease. PMID:29053735

Characterization of the first knock-out aldh7a1 zebrafish model for pyridoxine-dependent epilepsy using CRISPR-Cas9 technology.

PubMed

Zabinyakov, Nikita; Bullivant, Garrett; Cao, Feng; Fernandez Ojeda, Matilde; Jia, Zheng Ping; Wen, Xiao-Yan; Dowling, James J; Salomons, Gajja S; Mercimek-Andrews, Saadet

2017-01-01

Pyridoxine dependent epilepsy (PDE) is caused by likely pathogenic variants in ALDH7A1 (PDE-ALDH7A1) and inherited autosomal recessively. Neurotoxic alpha-amino adipic semialdehyde (alpha-AASA), piperideine 6-carboxylate and pipecolic acid accumulate in body fluids. Neonatal or infantile onset seizures refractory to anti-epileptic medications are clinical features. Treatment with pyridoxine, arginine and lysine-restricted diet does not normalize neurodevelopmental outcome or accumulation of neurotoxic metabolites. There is no animal model for high throughput drug screening. For this reason, we developed and characterized the first knock-out aldh7a1 zebrafish model using CRISPR-Cas9 technology. Zebrafish aldh7a1 mutants were generated by using a vector free method of CRISPR-Cas9 mutagenesis. Genotype analysis of aldh7a1 knock-out zebrafish was performed by high resolution melt analysis, direct sequencing and QIAxcel system. Electroencephalogram was performed. Alpha-AASA, piperideine 6-carboxylate and pipecolic acid, were measured by liquid chromatography-tandem mass spectrometry. Our knock-out aldh7a1 zebrafish has homozygous 5 base pair (bp) mutation in ALDH7A1. Knock-out aldh7a1 embryos have spontaneous rapid increase in locomotion and a rapid circling swim behavior earliest 8-day post fertilization (dpf). Electroencephalogram revealed large amplitude spike discharges compared to wild type. Knock-out aldh7a1 embryos have elevated alpha-AASA, piperideine 6-carboxylate and pipecolic acid compared to wild type embryos at 3 dpf. Knock-out aldh7a1 embryos showed no aldh7a1 protein by western blot compared to wild type. Our knock-out aldh7a1 zebrafish is a well characterized model for large-scale drug screening using behavioral and biochemical features and accurately recapitulates the human PDE-ALDH7A1 disease.

When Opportunity Knocks Twice: Dual Living Kidney Donation, Autonomy and the Public Interest

PubMed Central

Huxtable, Richard

2015-01-01

Abstract Living kidney transplantation offers the best treatment in terms of life‐expectancy and quality of life for those with end‐stage renal disease. The long‐term risks of living donor nephrectomy, although real, are very small, with evidence of good medium‐term outcomes. Who should be entitled to donate, and in which circumstances, is nevertheless a live question. We explore the ethical dimensions of a request by an individual to donate both of their kidneys during life: ‘dual living kidney donation’. Our ethical analysis is tethered to a hypothetical case study in which a father asks to donate a kidney to each of his twin boys. We explore the autonomy of the protagonists, alongside different dimensions of the public interest, such as the need to protect not only the recipients, but also the donor and even the wider community. Whilst acknowledging objections to ‘dual‐donation’, not least by reference to the harms that the donor might be expected to endure, we suggest there is a prima facie case for permitting this, provided that both donor and recipients are willing and that due attention is paid to such considerations as the autonomy and welfare of all parties, as well as to the wider ramifications of acting on such a request. We argue for broader interpretations of the concepts of autonomy and welfare, recognizing the importance of relationships and the relevance of more than merely physical well‐being. Equipped with such a holistic assessment, we suggest there is a prima facie case for allowing ‘dual living kidney donation’. PMID:26194324

An Analysis of Five Years of the Education Improvement Act on the Fiscal Capacity of South Carolina Schools To Deliver Equal Education Opportunity.

ERIC Educational Resources Information Center

Flanigan, J. L.; Richardson, M. D.

The first 5 years of South Carolina's 1984 Educational Improvement Act (EIA), a major educational reform program to deliver equal education opportunity and improve academic performance, are analyzed in this paper. A review of South Carolina's per-capita disposable income as compared with other states reveals an improvement in South Carolina's rank…

COLORADO STATE PLAN FOR ADULT BASIC EDUCATION UNDER PROVISIONS OF ADULT EDUCATION ACT OF 1966 (PL 89-750, NOV 3, 1966).

ERIC Educational Resources Information Center

Colorado State Dept. of Education, Denver.

LACK OF FINANCES, QUALIFIED PERSONNEL, AND TRAINING OPPORTUNITIES IN INSTITUTIONS OF HIGHER LEARNING MADE THE PROBLEMS OF EDUCATING THE 250,000 COLORADO ADULTS WITH LESS THAN EIGHT YEARS OF FORMAL SCHOOLING DIFFICULT TO ATTACK. UNDER THE ECONOMIC OPPORTUNITY ACT OF 1964, A COOPERATIVE EFFORT WAS BEGUN BY SCHOOL DISTRICTS AND OTHER AGENCIES TO…

77 FR 69933 - Expanding the Economic and Innovation Opportunities of Spectrum Through Incentive Auctions

Federal Register 2010, 2011, 2012, 2013, 2014

2012-11-21

...In the Notice of Proposed Rulemaking, ``Expanding the Economic and Innovation Opportunities of Spectrum Through Incentive Auctions'' (NPRM), released October 2, 2012, the Commission considers matters related to the implementation of Congress's mandate to conduct an incentive auction of broadcast television spectrum as set forth in the Middle Class Tax Relief and Job Creation Act of 2012 (Spectrum Act).

Healthy, Hunger-Free Kids Act of 2010: An Opportunity for School Nurses to Make a Difference

ERIC Educational Resources Information Center

Prokop, Jessica L.; Galon, Patricia

2011-01-01

Implementation of the Healthy Hunger-Free Kids Act of 2010 will provide an opportunity for school nurses to intervene in the serious childhood obesity problem in the United States. Major changes in the management of the National School Lunch Program (NSLP) will likely challenge schools yet may provide the impetus for a collaborative effort by the…

75 FR 60482 - Proposed Extension of Information Collection Request Submitted for Public Comment; Affordable...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-09-30

... Collection Request Submitted for Public Comment; Affordable Care Act Enrollment Opportunity Notice Relating to Dependent Coverage; Affordable Care Act Grandfathered Health Plan Disclosure and Recordkeeping Requirement; Affordable Care Act Rescission Notice; Affordable Care Act Patient Protections Notice; Affordable...

16 CFR 309.17 - Labels.

Code of Federal Regulations, 2010 CFR

2010-01-01

... height) knocked out of a 1″ (2.54 cm) deep band. The type for the words “MINIMUM” and the principal... should be set as “normal.” The type for the fuel name is 50 point (1/2″ 1.27 cm) cap height) knocked out... point (1/2″ 1.27 cm) cap height) knocked out of a 1″ (2.54 cm) deep band. All other type is 24 pt. (1/4...

16 CFR 309.17 - Labels.

Code of Federal Regulations, 2011 CFR

2011-01-01

... height) knocked out of a 1″ (2.54 cm) deep band. The type for the words “MINIMUM” and the principal... should be set as “normal.” The type for the fuel name is 50 point (1/2″ 1.27 cm) cap height) knocked out... point (1/2″ 1.27 cm) cap height) knocked out of a 1″ (2.54 cm) deep band. All other type is 24 pt. (1/4...

Targeting Transcription Elongation Machinery for Breast Cancer Therapy

DTIC Science & Technology

2017-05-01

be performed to evaluate the pausing index for RNA Pol II. The potential role of a Super Enhancer will also be tested by knocking down the mediator...generation of all the cell lines stably knocking out the components of various P-TEFb complexes and performed some of the biochemical experiments...assessing the changes in P-TEFb complex formation upon knocking down or overexpression of various components. Funding Support: NIH Has there been a

Targeting the Prometastatic Microenvironment of the Involuting mammary gland

DTIC Science & Technology

2016-09-01

metastatic variants and by lentiviral knock down. 15. SUBJECT TERMS Cell Adhesion, Involution, Metastasis, Latent TGFbeta Binding Protein, Pregnancy...metastatic parental cell line (Report year 2 Fig. 4). We generated a lentiviral knock down system that effectively reduced LTBP1 expression within this cell...this pilot set of animals and will be infecting and following a larger cohort by IVIS to track the effect of knocking down Ltbp1 on metastasis in

Efficient gene knock-out and knock-in with transgenic Cas9 in Drosophila.

PubMed

Xue, Zhaoyu; Ren, Mengda; Wu, Menghua; Dai, Junbiao; Rong, Yikang S; Gao, Guanjun

2014-03-21

Bacterial Cas9 nuclease induces site-specific DNA breaks using small gRNA as guides. Cas9 has been successfully introduced into Drosophila for genome editing. Here, we improve the versatility of this method by developing a transgenic system that expresses Cas9 in the Drosophila germline. Using this system, we induced inheritable knock-out mutations by injecting only the gRNA into embryos, achieved highly efficient mutagenesis by expressing gRNA from the promoter of a novel non-coding RNA gene, and recovered homologous recombination-based knock-in of a fluorescent marker at a rate of 4.5% by co-injecting gRNA with a circular DNA donor. Copyright © 2014 Xue et al.

24 CFR 5.105 - Other Federal requirements.

Code of Federal Regulations, 2010 CFR

2010-04-01

... the respective program regulations: (a) Nondiscrimination and equal opportunity. The Fair Housing Act... Opportunity in Housing Programs) and implementing regulations at 24 CFR part 107; title VI of the Civil Rights...) (Equal Employment Opportunity Programs) and implementing regulations at 41 CFR chapter 60; Executive...

The caspase-3/p120 RasGAP stress-sensing module reduces liver cancer incidence but does not affect overall survival in gamma-irradiated and carcinogen-treated mice.

PubMed

Vanli, Güliz; Sempoux, Christine; Widmann, Christian

2017-06-01

Activation of oncogenes is the initial step in cellular transformation. Oncogenes favor aberrant proliferation, which, at least initially, induces cellular stress. This oncogenic stress can act as a safeguard mechanism against further transformation by inducing senescence or apoptosis. Yet, the few premalignant cells that tolerate and escape these senescent or apoptotic responses are those that will ultimately generate tumors. The caspase-3/p120 RasGAP module is a stress-sensing device that promotes survival under mild stress conditions. A point mutation in RasGAP that prevents its cleavage by caspase-3 inactivates the pro-survival capacity of the device. When the mice homozygous for this mutation (D455A knock-in mice) are patho-physiologically challenged, they experience much stronger cellular damage than their wild-type counterparts and the affected organs rapidly lose their functionality. We reasoned that the caspase-3/p120 RasGAP module could help premalignant cells to cope with oncogenic stress and hence favor the development of tumors. Using gamma-irradiation and N-ethyl-N-nitrosourea (ENU) as tumor initiators, we assessed the survival advantage that the caspase-3/p120 RasGAP module could provide to premalignant cells. No difference in overall mortality between wild-type and D455A knock-in mice were observed. However, the number of ENU-induced liver tumors in the knock-in mice was higher than in control mice. These results indicate that the caspase-3/p120 RasGAP stress-sensing module impacts on carcinogen-induced liver cancer incidence but not sufficiently so as to affect overall survival. Hence, gamma irradiation and ENU-induced tumorigenesis processes do not critically rely on a survival mechanism that contributes to the maintenance of organ homeostasis in stressed healthy tissues. © 2017 Wiley Periodicals, Inc.

The Changing Face of Immigration Law.

ERIC Educational Resources Information Center

Nugent, Christopher

2001-01-01

Focuses on laws that influence U.S. immigration, such as the Illegal Immigration Reform and Immigrant Responsibility Act (1996), the Personal Responsibility and Work Opportunity Reconciliation Act (1996), the Anti-Terrorism and Effective Death Penalty Act (1996), and the Legal Immigration and Family Equity Act (2000). Includes discussion…

Seizing opportunities under the Affordable Care Act for transforming the mental and behavioral health system.

PubMed

Mechanic, David

2012-02-01

The Affordable Care Act, along with Medicaid expansions, offers the opportunity to redesign the nation's highly flawed mental health system. It promotes new programs and tools, such as health homes, interdisciplinary care teams, the broadening of the Medicaid Home and Community-Based Services option, co-location of physical health and behavioral services, and collaborative care. Provisions of the act offer extraordinary opportunities, for instance, to insure many more people, reimburse previously unreimbursed services, integrate care using new information technology tools and treatment teams, confront complex chronic comorbidities, and adopt underused evidence-based interventions. The Centers for Medicare and Medicaid Services and its Center for Medicare and Medicaid Innovation should work intensively with the states to implement these new programs and other arrangements and begin to fulfill the many unmet promises of community mental health care.

Glutaminyl Cyclase Knock-out Mice Exhibit Slight Hypothyroidism but No Hypogonadism

PubMed Central

Schilling, Stephan; Kohlmann, Stephanie; Bäuscher, Christoph; Sedlmeier, Reinhard; Koch, Birgit; Eichentopf, Rico; Becker, Andreas; Cynis, Holger; Hoffmann, Torsten; Berg, Sabine; Freyse, Ernst-Joachim; von Hörsten, Stephan; Rossner, Steffen; Graubner, Sigrid; Demuth, Hans-Ulrich

2011-01-01

Glutaminyl cyclases (QCs) catalyze the formation of pyroglutamate (pGlu) residues at the N terminus of peptides and proteins. Hypothalamic pGlu hormones, such as thyrotropin-releasing hormone and gonadotropin-releasing hormone are essential for regulation of metabolism and fertility in the hypothalamic pituitary thyroid and gonadal axes, respectively. Here, we analyzed the consequences of constitutive genetic QC ablation on endocrine functions and on the behavior of adult mice. Adult homozygous QC knock-out mice are fertile and behave indistinguishably from wild type mice in tests of motor function, cognition, general activity, and ingestion behavior. The QC knock-out results in a dramatic drop of enzyme activity in the brain, especially in hypothalamus and in plasma. Other peripheral organs like liver and spleen still contain QC activity, which is most likely caused by its homolog isoQC. The serum gonadotropin-releasing hormone, TSH, and testosterone concentrations were not changed by QC depletion. The serum thyroxine was decreased by 24% in homozygous QC knock-out animals, suggesting a mild hypothyroidism. QC knock-out mice were indistinguishable from wild type with regard to blood glucose and glucose tolerance, thus differing from reports of thyrotropin-releasing hormone knock-out mice significantly. The results suggest a significant formation of the hypothalamic pGlu hormones by alternative mechanisms, like spontaneous cyclization or conversion by isoQC. The different effects of QC depletion on the hypothalamic pituitary thyroid and gonadal axes might indicate slightly different modes of substrate conversion of both enzymes. The absence of significant abnormalities in QC knock-out mice suggests the presence of a therapeutic window for suppression of QC activity in current drug development. PMID:21330373

Hyperactivity of newborn Pten knock-out neurons results from increased excitatory synaptic drive.

PubMed

Williams, Michael R; DeSpenza, Tyrone; Li, Meijie; Gulledge, Allan T; Luikart, Bryan W

2015-01-21

Developing neurons must regulate morphology, intrinsic excitability, and synaptogenesis to form neural circuits. When these processes go awry, disorders, including autism spectrum disorder (ASD) or epilepsy, may result. The phosphatase Pten is mutated in some patients having ASD and seizures, suggesting that its mutation disrupts neurological function in part through increasing neuronal activity. Supporting this idea, neuronal knock-out of Pten in mice can cause macrocephaly, behavioral changes similar to ASD, and seizures. However, the mechanisms through which excitability is enhanced following Pten depletion are unclear. Previous studies have separately shown that Pten-depleted neurons can drive seizures, receive elevated excitatory synaptic input, and have abnormal dendrites. We therefore tested the hypothesis that developing Pten-depleted neurons are hyperactive due to increased excitatory synaptogenesis using electrophysiology, calcium imaging, morphological analyses, and modeling. This was accomplished by coinjecting retroviruses to either "birthdate" or birthdate and knock-out Pten in granule neurons of the murine neonatal dentate gyrus. We found that Pten knock-out neurons, despite a rapid onset of hypertrophy, were more active in vivo. Pten knock-out neurons fired at more hyperpolarized membrane potentials, displayed greater peak spike rates, and were more sensitive to depolarizing synaptic input. The increased sensitivity of Pten knock-out neurons was due, in part, to a higher density of synapses located more proximal to the soma. We determined that increased synaptic drive was sufficient to drive hypertrophic Pten knock-out neurons beyond their altered action potential threshold. Thus, our work contributes a developmental mechanism for the increased activity of Pten-depleted neurons. Copyright © 2015 the authors 0270-6474/15/350943-17$15.00/0.

Generation of Knock-in Mouse by Genome Editing.

PubMed

Fujii, Wataru

2017-01-01

Knock-in mice are useful for evaluating endogenous gene expressions and functions in vivo. Instead of the conventional gene-targeting method using embryonic stem cells, an exogenous DNA sequence can be inserted into the target locus in the zygote using genome editing technology. In this chapter, I describe the generation of epitope-tagged mice using engineered endonuclease and single-stranded oligodeoxynucleotide through the mouse zygote as an example of how to generate a knock-in mouse by genome editing.

Identification of rat Rosa26 locus enables generation of knock-in rat lines ubiquitously expressing tdTomato.

PubMed

Kobayashi, Toshihiro; Kato-Itoh, Megumi; Yamaguchi, Tomoyuki; Tamura, Chihiro; Sanbo, Makoto; Hirabayashi, Masumi; Nakauchi, Hiromitsu

2012-11-01

Recent discovery of a method for derivation and culture of germline-competent rat pluripotent stem cells (PSCs) enables generation of transgenic rats or knock-out rats via genetic modification of such PSCs. This opens the way to use rats, as is routine in mice, for analyses of gene functions or physiological features. In mouse or human, one widely used technique to express a gene of interest stably and ubiquitously is to insert that gene into the Rosa26 locus via gene targeting of PSCs. Rosa26 knock-in mice conditionally expressing a reporter or a toxin gene have contributed to tracing or ablation of specific cell lineages. We successfully identified a rat orthologue of the mouse Rosa26 locus. Insertion of tdTomato, a variant of red fluorescent protein, into the Rosa26 locus of PSCs of various rat strains allows ubiquitous expression of tdTomato. Through germline transmission of one Rosa26-tdTomato knock-in embryonic stem cell line, we also obtained tdTomato knock-in rats. These expressed tdTomato ubiquitously throughout their bodies, which indicates that the rat Rosa26 locus conserves functions of its orthologues in mouse and human. The new tools described here (targeting vectors, knock-in PSCs, and rats) should be useful for a variety of research using rats.

Promoting Prevention Through the Affordable Care Act: Workplace Wellness

PubMed Central

Roffenbender, Jason S.; Goetzel, Ron Z.; Millard, Francois; Wildenhaus, Kevin; DeSantis, Charles; Novelli, William

2012-01-01

Public health in the United States can be improved by building workplace “cultures of health” that support healthy lifestyles. The Affordable Care Act (ACA), which includes the Prevention and Public Health Fund, will support a new focus on prevention and wellness, offering opportunities to strengthen the public’s health through workplace wellness initiatives. This article describes the opportunity the ACA provides to improve worker wellness. PMID:23237245

Helping Employers Comply with the ADA: An Assessment of How the United States Equal Employment Opportunity Commission Is Enforcing Title I of the Americans with Disabilities Act.

ERIC Educational Resources Information Center

Isler, Frederick D.; Zalokar, Nadja; Chambers, David; Kraus, Rebecca; Johnson, Wanda; Butler, Margaret; Avery, Michelle Leigh; Tyler, Marcia; Baird, Andrea; Foshee, Latrice; Turner, Ilona

This report from the United States Commission on Civil Rights focuses specifically on the efforts of the U.S. Equal Employment Opportunities Commission (EEOC) to enforce Title I of the Americans with Disabilities Act, which prohibits discrimination based on disability in employment. The report evaluates and analyzes EEOC's regulations and policies…

Efficient generation of Rosa26 knock-in mice using CRISPR/Cas9 in C57BL/6 zygotes.

PubMed

Chu, Van Trung; Weber, Timm; Graf, Robin; Sommermann, Thomas; Petsch, Kerstin; Sack, Ulrike; Volchkov, Pavel; Rajewsky, Klaus; Kühn, Ralf

2016-01-16

The CRISPR/Cas9 system is increasingly used for gene inactivation in mouse zygotes, but homology-directed mutagenesis and use of inbred embryos are less established. In particular, Rosa26 knock-in alleles for the insertion of transgenes in a genomic 'safe harbor' site, have not been produced. Here we applied CRISPR/Cas9 for the knock-in of 8-11 kb inserts into Rosa26 of C57BL/6 zygotes. We found that 10-20 % of live pups derived from microinjected zygotes were founder mutants, without apparent off-target effects, and up to 50 % knock-in embryos were recovered upon coinjection of Cas9 mRNA and protein. Using this approach, we established a new mouse line for the Cre/loxP-dependent expression of Cas9. Altogether, our protocols and resources support the fast and direct generation of new Rosa26 knock-in alleles and of Cas9-mediated in vivo gene editing in the widely used C57BL/6 inbred strain.

Targeted knock-in of an scFv-Fc antibody gene into the hprt locus of Chinese hamster ovary cells using CRISPR/Cas9 and CRIS-PITCh systems.

PubMed

Kawabe, Yoshinori; Komatsu, Shinya; Komatsu, Shodai; Murakami, Mai; Ito, Akira; Sakuma, Tetsushi; Nakamura, Takahiro; Yamamoto, Takashi; Kamihira, Masamichi

2018-05-01

Chinese hamster ovary (CHO) cells have been used as host cells for the production of pharmaceutical proteins. For the high and stable production of target proteins, the transgene should be integrated into a suitable genomic locus of host cells. Here, we generated knock-in CHO cells, in which transgene cassettes without a vector backbone sequence were integrated into the hprt locus of the CHO genome using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 and CRISPR-mediated precise integration into target chromosome (CRIS-PITCh) systems. We investigated the efficiency of targeted knock-in of transgenes using these systems. As a practical example, we generated knock-in CHO cells producing an scFv-Fc antibody using the CRIS-PITCh system mediated by microhomology sequences for targeting. We found that the CRIS-PITCh system can facilitate targeted knock-in for CHO cell engineering. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

78 FR 44949 - Agency Information Collection Activities: Submission for OMB Review; Final Comment Request

Federal Register 2010, 2011, 2012, 2013, 2014

2013-07-25

... EQUAL EMPLOYMENT OPPORTUNITY COMMISSION Agency Information Collection Activities: Submission for OMB Review; Final Comment Request AGENCY: Equal Employment Opportunity Commission. ACTION: Final... Reduction Act of 1995, the Equal Employment Opportunity Commission (Commission or EEOC) gives notice that it...

77 FR 43498 - Federal Sector Equal Employment Opportunity

Federal Register 2010, 2011, 2012, 2013, 2014

2012-07-25

... on the basis of age; the Equal Pay Act of 1963, which prohibits sex-based wage discrimination; and..., Age discrimination, Equal employment opportunity, Government employees, Individuals with disabilities... EQUAL EMPLOYMENT OPPORTUNITY COMMISSION 29 CFR Part 1614 RIN Number 3046-AA73 Federal Sector Equal...

75 FR 70342 - Privacy Act; System of Records: Equal Employment Opportunity Records

Federal Register 2010, 2011, 2012, 2013, 2014

2010-11-17

... users are given cyber security awareness training which covers the procedures for handling Sensitive but.... State-09 SYSTEM NAME: Equal Employment Opportunity Records. SECURITY CLASSIFICATION: Unclassified... apply to the Equal Employment Opportunity Records, State-09. DISCLOSURE TO CONSUMER REPORTING AGENCIES...

29 CFR 1625.2 - Discrimination prohibited by the Act.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 29 Labor 4 2012-07-01 2012-07-01 false Discrimination prohibited by the Act. 1625.2 Section 1625.2 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION AGE DISCRIMINATION IN EMPLOYMENT ACT Interpretations § 1625.2 Discrimination prohibited by the Act. It is unlawful...

29 CFR 1625.2 - Discrimination prohibited by the Act.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 29 Labor 4 2013-07-01 2013-07-01 false Discrimination prohibited by the Act. 1625.2 Section 1625.2 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION AGE DISCRIMINATION IN EMPLOYMENT ACT Interpretations § 1625.2 Discrimination prohibited by the Act. It is unlawful...

29 CFR 1625.2 - Discrimination prohibited by the Act.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 29 Labor 4 2014-07-01 2014-07-01 false Discrimination prohibited by the Act. 1625.2 Section 1625.2 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION AGE DISCRIMINATION IN EMPLOYMENT ACT Interpretations § 1625.2 Discrimination prohibited by the Act. It is unlawful...

29 CFR 1625.2 - Discrimination prohibited by the Act.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 29 Labor 4 2011-07-01 2011-07-01 false Discrimination prohibited by the Act. 1625.2 Section 1625.2 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION AGE DISCRIMINATION IN EMPLOYMENT ACT Interpretations § 1625.2 Discrimination prohibited by the Act. It is unlawful...

29 CFR 1625.2 - Discrimination prohibited by the Act.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 29 Labor 4 2010-07-01 2010-07-01 false Discrimination prohibited by the Act. 1625.2 Section 1625.2 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION AGE DISCRIMINATION IN EMPLOYMENT ACT Interpretations § 1625.2 Discrimination prohibited by the Act. It is unlawful...

Secretarial Administraton: The Interviewing Process.

ERIC Educational Resources Information Center

Nemesh, Anna

1979-01-01

Suggests classroom techniques to prepare business students for employment interviews and gives information on lawful and unlawful employment interview inquiries, as well as some fair employment legal requirements of the Equal Employment Opportunity Act of 1974, Civil Rights Act of 1964, Equal Pay Act of 1963, and Rehabilitation Act of 1973. (MF)

12 CFR 268.202 - Equal Pay Act.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 12 Banks and Banking 3 2010-01-01 2010-01-01 false Equal Pay Act. 268.202 Section 268.202 Banks... REGARDING EQUAL OPPORTUNITY Provisions Applicable to Particular Complaints § 268.202 Equal Pay Act. Complaints alleging violations of the Equal Pay Act shall be processed under this part. ...

Differential Effector Engagement by Oncogenic KRAS.

PubMed

Yuan, Tina L; Amzallag, Arnaud; Bagni, Rachel; Yi, Ming; Afghani, Shervin; Burgan, William; Fer, Nicole; Strathern, Leslie A; Powell, Katie; Smith, Brian; Waters, Andrew M; Drubin, David; Thomson, Ty; Liao, Rosy; Greninger, Patricia; Stein, Giovanna T; Murchie, Ellen; Cortez, Eliane; Egan, Regina K; Procter, Lauren; Bess, Matthew; Cheng, Kwong Tai; Lee, Chih-Shia; Lee, Liam Changwoo; Fellmann, Christof; Stephens, Robert; Luo, Ji; Lowe, Scott W; Benes, Cyril H; McCormick, Frank

2018-02-13

KRAS can bind numerous effector proteins, which activate different downstream signaling events. The best known are RAF, phosphatidylinositide (PI)-3' kinase, and RalGDS families, but many additional direct and indirect effectors have been reported. We have assessed how these effectors contribute to several major phenotypes in a quantitative way, using an arrayed combinatorial siRNA screen in which we knocked down 41 KRAS effectors nodes in 92 cell lines. We show that every cell line has a unique combination of effector dependencies, but in spite of this heterogeneity, we were able to identify two major subtypes of KRAS mutant cancers of the lung, pancreas, and large intestine, which reflect different KRAS effector engagement and opportunities for therapeutic intervention. Copyright © 2018 The Author(s). Published by Elsevier Inc. All rights reserved.

Turning K-12 Science Education Inside Out, Knocking Down Walls and Empowering the Disenchanted.

NASA Astrophysics Data System (ADS)

Lin, A. Y. M.

2016-12-01

For a 'user' there are several genres of citizen science activities one can enlist themselves in, from microtasked analytics to data collection. Often times design conversation for these efforts are focused around the goal of collecting high quality data for an urgent scientific question. However, there is much to be discussed around the opportunity to expand upon the interaction experience of the 'user'. This is particularly relevant in the integration of citizen science in the classroom. Here we explore the role of citizen science in formal K-12 science education through the lens of "Project Based Learning", examining design challenges in classroom adoption (including standards alignment) as well as interaction design focused around long term user/student motivation and engagement in the science exploration.

A Genome-Wide Knockout Screen to Identify Genes Involved in Acquired Carboplatin Resistance

DTIC Science & Technology

2016-07-01

library screen to identify genes that when knocked out render human ovarian cells > 2.5-fold resistant to CBDCA; 2) Validate the ability of...a GeCKOv2 library screen to identify genes that when knocked out render human ovarian cells > 2.5-fold resistant to CBDCA; 2) validate the ability of...resistance in either cell lines or clinical samples. The CRIPSR-cas9 technology now provides us with a major new tool to introduce knock out mutations

Cancer-associated Isocitrate Dehydrogenase 1 (IDH1) R132H Mutation and d-2-Hydroxyglutarate Stimulate Glutamine Metabolism under Hypoxia*

PubMed Central

Reitman, Zachary J.; Duncan, Christopher G.; Poteet, Ethan; Winters, Ali; Yan, Liang-Jun; Gooden, David M.; Spasojevic, Ivan; Boros, Laszlo G.; Yang, Shao-Hua; Yan, Hai

2014-01-01

Mutations in the cytosolic NADP+-dependent isocitrate dehydrogenase (IDH1) occur in several types of cancer, and altered cellular metabolism associated with IDH1 mutations presents unique therapeutic opportunities. By altering IDH1, these mutations target a critical step in reductive glutamine metabolism, the metabolic pathway that converts glutamine ultimately to acetyl-CoA for biosynthetic processes. While IDH1-mutated cells are sensitive to therapies that target glutamine metabolism, the effect of IDH1 mutations on reductive glutamine metabolism remains poorly understood. To explore this issue, we investigated the effect of a knock-in, single-codon IDH1-R132H mutation on the metabolism of the HCT116 colorectal adenocarcinoma cell line. Here we report the R132H-isobolome by using targeted 13C isotopomer tracer fate analysis to trace the metabolic fate of glucose and glutamine in this system. We show that introduction of the R132H mutation into IDH1 up-regulates the contribution of glutamine to lipogenesis in hypoxia, but not in normoxia. Treatment of cells with a d-2-hydroxyglutarate (d-2HG) ester recapitulated these changes, indicating that the alterations observed in the knocked-in cells were mediated by d-2HG produced by the IDH1 mutant. These studies provide a dynamic mechanistic basis for metabolic alterations observed in IDH1-mutated tumors and uncover potential therapeutic targets in IDH1-mutated cancers. PMID:24986863

75 FR 56057 - Proposed Information Collection; Comment Request; Broadband Technology Opportunities Program Post...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-09-15

... Information Collection; Comment Request; Broadband Technology Opportunities Program Post-Award Quarterly and... (Recovery Act) establishes and provides $4.7 billion for the Broadband Technology Opportunities Program... million will be made available for competitive grants to expand public computer center capacity; at least...

Absence of ERRα in Female Mice Confers Resistance to Bone Loss Induced by Age or Estrogen-Deficiency

PubMed Central

Rabier, Bénédicte; Monfoulet, Laurent; Dine, Julien; Macari, Claire; Espallergues, Julie; Horard, Béatrice; Giguère, Vincent; Cohen-Solal, Martine; Chassande, Olivier; Vanacker, Jean-Marc

2009-01-01

Background ERRα is an orphan member of the nuclear hormone receptor superfamily, which acts as a transcription factor and is involved in various metabolic processes. ERRα is also highly expressed in ossification zones during mouse development as well as in human bones and cell lines. Previous data have shown that this receptor up-modulates the expression of osteopontin, which acts as an inhibitor of bone mineralization and whose absence results in resistance to ovariectomy-induced bone loss. Altogether this suggests that ERRα may negatively regulate bone mass and could impact on bone fragility that occurs in the absence of estrogens. Methods/Principal Findings In this report, we have determined the in vivo effect of ERRα on bone, using knock-out mice. Relative to wild type animals, female ERRαKO bones do not age and are resistant to bone loss induced by estrogen-withdrawal. Strikingly male ERRαKO mice are indistinguishable from their wild type counterparts, both at the unchallenged or gonadectomized state. Using primary cell cultures originating from ERRαKO bone marrow, we also show that ERRα acts as an inhibitor of osteoblast differentiation. Conclusion/Significance Down-regulating ERRα could thus be beneficial against osteoporosis. PMID:19936213

Mapping Heart Development in Flies: Src42A Acts Non-Autonomously to Promote Heart Tube Formation in Drosophila

PubMed Central

Vanderploeg, Jessica; Jacobs, J. Roger

2017-01-01

Congenital heart defects, clinically identified in both small and large animals, are multifactorial and complex. Although heritable factors are known to have a role in cardiovascular disease, the full genetic aetiology remains unclear. Model organism research has proven valuable in providing a deeper understanding of the essential factors in heart development. For example, mouse knock-out studies reveal a role for the Integrin adhesion receptor in cardiac tissue. Recent research in Drosophila melanogaster (the fruit fly), a powerful experimental model, has demonstrated that the link between the extracellular matrix and the cell, mediated by Integrins, is required for multiple aspects of cardiogenesis. Here we test the hypothesis that Integrins signal to the heart cells through Src42A kinase. Using the powerful genetics and cell biology analysis possible in Drosophila, we demonstrate that Src42A acts in early events of heart tube development. Careful examination of mutant heart tissue and genetic interaction data suggests that Src42A’s role is independent of Integrin and the Integrin-related Focal Adhesion Kinase. Rather, Src42A acts non-autonomously by promoting programmed cell death of the amnioserosa, a transient tissue that neighbors the developing heart. PMID:29056682

Genetics Home Reference: mucopolysaccharidosis type IV

MedlinePlus

... individuals develop various skeletal abnormalities, including short stature, knock knees, and abnormalities of the ribs, chest, spine, ... links) Encyclopedia: Cloudy cornea Encyclopedia: Hypermobile joints Encyclopedia: Knock ... Morquio syndrome Encyclopedia: Mucopolysaccharides Health Topic: ...

Lack of huntingtin promotes neural stem cells differentiation into glial cells while neurons expressing huntingtin with expanded polyglutamine tracts undergo cell death.

PubMed

Conforti, Paola; Camnasio, Stefano; Mutti, Cesare; Valenza, Marta; Thompson, Morgan; Fossale, Elisa; Zeitlin, Scott; MacDonald, Marcy E; Zuccato, Chiara; Cattaneo, Elena

2013-02-01

Huntington's disease (HD) is a neurodegenerative disorder that affects muscle coordination and diminishes cognitive abilities. The genetic basis of the disease is an expansion of CAG repeats in the Huntingtin (Htt) gene. Here we aimed to generate a series of mouse neural stem (NS) cell lines that carried varying numbers of CAG repeats in the mouse Htt gene (Hdh CAG knock-in NS cells) or that had Hdh null alleles (Hdh knock-out NS cells). Towards this end, Hdh CAG knock-in mouse ES cell lines that carried an Htt gene with 20, 50, 111, or 140 CAG repeats or that were Htt null were neuralized and converted into self-renewing NS cells. The resulting NS cell lines were immunopositive for the neural stem cell markers NESTIN, SOX2, and BLBP and had similar proliferative rates and cell cycle distributions. After 14 days in vitro, wild-type NS cells gave rise to cultures composed of 70% MAP2(+) neurons and 30% GFAP(+) astrocytes. In contrast, NS cells with expanded CAG repeats underwent neuronal cell death, with only 38%±15% of the MAP2(+) cells remaining at the end of the differentiation period. Cell death was verified by increased caspase 3/7 activity on day 14 of the neuronal differentiation protocol. Interestingly, Hdh knock-out NS cells treated using the same neuronal differentiation protocol showed a dramatic increase in the number of GFAP(+) cells on day 14 (61%±20% versus 24%±10% in controls), and a massive decrease of MAP2(+) neurons (30%±11% versus 64%±17% in controls). Both Hdh CAG knock-in NS cells and Hdh knock-out NS cells showed reduced levels of Bdnf mRNA during neuronal differentiation, in agreement with data obtained previously in HD mouse models and in post-mortem brain samples from HD patients. We concluded that Hdh CAG knock-in and Hdh knock-out NS cells have potential as tools for investigating the roles of normal and mutant HTT in differentiated neurons and glial cells of the brain. Copyright © 2012 Elsevier Inc. All rights reserved.

The Integration of Minorities into Special Operations: How Cultural Diversity Enhances Operations

DTIC Science & Technology

2014-03-01

served George Washington’s Continental Army beginning in 1775. Then, it was about capability and concerns over ethnic and gender equality , while... equal opportunity than capability. In 2009, a Diversity Commission established by the United States Government, argued that the ranks of the...such as Title VII of the Civil Rights Act of 1964 and the Equal Employment Opportunity Act of 1972, have aided the country in moving past

Evaluation of the DC Opportunity Scholarship Program: An Early Look at Applicants and Participating Schools under the SOAR Act. Year 1 Report. NCEE 2015-4000

ERIC Educational Resources Information Center

Feldman, Jill; Lucas-McLean, Juanita; Gutmann, Babette; Dynarski, Mark; Betts, Julian

2014-01-01

This report explores implementation of the District of Columbia Opportunity Scholarship Program (OSP) in the first two years after Congress reauthorized it with some changes under the SOAR Act of 2011. Key findings include the following: (1) Just over half of all DC private schools participated in the OSP, with current schools more likely to have…

Evaluation of the DC Opportunity Scholarship Program: An Early Look at Applicants and Participating Schools under the SOAR Act. Year 1 Report, Executive Summary. NCEE 2015-4000

ERIC Educational Resources Information Center

Feldman, Jill; Lucas-McLean, Juanita; Gutmann, Babette; Dynarski, Mark; Betts, Julian

2014-01-01

This report explores implementation of the District of Columbia Opportunity Scholarship Program (OSP) in the first two years after Congress reauthorized it with some changes under the SOAR Act of 2011. Key findings include the following: (1) Just over half of all DC private schools participated in the OSP, with current schools more likely to have…

Examining Opportunity-to-Learn and Success in High School Mathematics Performance in California under NCLB

ERIC Educational Resources Information Center

Gavrilovic, Daniel Miodrag

2013-01-01

The No Child Left Behind Act of 2001 has put many schools under a lot of pressure to meet its high demands. In this quantitative study, the effects that the NCLB act has had on students' opportunity to learn (OTL) and Subject Level Success (SS) from 2004 to 2012 in 9th, 10th, and 11th grade math coursework (Algebra 1, Geometry, Algebra 2, and…

Hawaiian Homeownership Opportunity Act of 2011

THOMAS, 112th Congress

Rep. Hirono, Mazie K. [D-HI-2

2011-07-26

House - 08/22/2011 Referred to the Subcommittee on Insurance, Housing and Community Opportunity. (All Actions) Tracker: This bill has the status IntroducedHere are the steps for Status of Legislation:

Still Not Equal: Expanding Educational Opportunity in Society

ERIC Educational Resources Information Center

Brown, M. Christopher, II, Ed.

2007-01-01

"Still Not Equal: Expanding Educational Opportunity in Society" addresses the successes and failures of "Brown v. Board of Education" and the Civil Rights Act of 1964, as well as the continuing challenge of expanding educational opportunity in the United States and across the Black diaspora. The educational, political, and social influence…

Students with Intellectual Disabilities Going to College? Absolutely!

ERIC Educational Resources Information Center

Kleinert, Harold L.; Jones, Melissa M.; Sheppard-Jones, Kathleen; Harp, Beverly; Harrison, Elizabeth M.

2012-01-01

In this article we describe the new opportunities for postsecondary education available to students with intellectual disabilities (ID) through the Higher Education Opportunities Act of 2008. We also identify the importance of these opportunities for students with ID, and key strategies for ensuring that their college experience is an important…

Contaminated land and wetland remediation in Nigeria: Opportunities for sustainable livelihood creation.

PubMed

Sam, K; Zabbey, N

2018-10-15

The Niger Delta region of Nigeria is one of the most crude oil impacted deltas globally. The region has experienced over five decades of oil related contamination of the total environment (air, soil, water and biota). In 2011, UNEP released a seminal report on oil impact on Ogoniland environments, which up scaled demands for urgent clean up and restoration of degraded bio-resource rich environments of the Niger Delta, starting from Ogoniland. The Nigerian Government demonstrated renewed political will to remediate contaminated sites in Ogoniland with a launch of the clean-up exercise in June 2016. Stakeholders' expectations from the clean-up include not only environmental remediation but also restoration and creation of sustainable livelihood opportunities to reduce poverty in the region. Most studies have focused on the environmental restoration aspect and identified bioremediation as the likely appropriate remediation approach for Ogoniland, given its low environmental footprints, and low-cost burden on the weak and overstretched economy of Nigeria. This study mapped opportunities for sustainable livelihood creation during the Ogoniland remediation and restoration exercise. Given the value chain of bioremediation and its ancillary activities, the study analysed opportunities and mechanisms for skilled and unskilled job creation and prospects for sustainable livelihoods and knock-on effects. It is anticipated that the clean-up process would lead to economic prosperity and mitigate resource-driven conflicts in the Niger Delta. The study provides an exemplar for waste-to-wealth transformation in regions where natural resource mining has impacted communities, and has dislocated local economies and age-old livelihood structures. Copyright © 2018. Published by Elsevier B.V.

32 CFR 505.3 - Privacy Act systems of records.

Code of Federal Regulations, 2010 CFR

2010-07-01

... AND PUBLIC RELATIONS ARMY PRIVACY ACT PROGRAM § 505.3 Privacy Act systems of records. (a) Systems of... Register in a system of records notice, which will provide the public an opportunity to comment before DA... Register on new, amended, altered, or deleted systems of records to inform the public of the Privacy Act...

Higher Education Opportunity Act (Public Law 110-315)

ERIC Educational Resources Information Center

US Congress, 2008

2008-01-01

This purpose of this Act is to amend and extend the Higher Education Act of 1965, and for other purposes. This Act includes the following titles: (1) General Provisions; (2) Teacher Quality Enhancement; (3) Institutional Aid; (4) Student Assistance (Grants to Students in Attendance at Institutions of Higher Education, Federal Family Education Loan…

29 CFR 1614.202 - Equal Pay Act.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 29 Labor 4 2010-07-01 2010-07-01 false Equal Pay Act. 1614.202 Section 1614.202 Labor Regulations... OPPORTUNITY Provisions Applicable to Particular Complaints § 1614.202 Equal Pay Act. (a) In its enforcement of the Equal Pay Act, the Commission has the authority to investigate an agency's employment practices on...

Genetics Home Reference: hereditary hypophosphatemic rickets

MedlinePlus

... noticeable of these abnormalities are bowed legs or knock knees (a condition in which the lower legs ... Information & Resources MedlinePlus (4 links) Encyclopedia: Bowlegs Encyclopedia: Knock Knees Encyclopedia: Rickets Health Topic: Rickets Genetic and ...

ARPP-16 Is a Striatal-Enriched Inhibitor of Protein Phosphatase 2A Regulated by Microtubule-Associated Serine/Threonine Kinase 3 (Mast 3 Kinase)

PubMed Central

Andrade, Erika C.; Musante, Veronica; Horiuchi, Atsuko; Greengard, Paul; Taylor, Jane R.

2017-01-01

ARPP-16 (cAMP-regulated phospho-protein of molecular weight 16 kDa) is one of several small acid-soluble proteins highly expressed in medium spiny neurons of striatum that are phosphorylated in response to dopamine acting via D1 receptor/protein kinase A (PKA) signaling. We show here that ARPP-16 is also phosphorylated in vitro and in vivo by microtubule-associated serine/threonine kinase 3 (MAST3 kinase), an enzyme of previously unknown function that is enriched in striatum. We find that ARPP-16 interacts directly with the scaffolding A subunit of the serine/threonine protein phosphatase, PP2A, and that phosphorylation of ARPP-16 at Ser46 by MAST3 kinase converts the protein into a selective inhibitor of B55α- and B56δ-containing heterotrimeric forms of PP2A. Ser46 of ARPP-16 is phosphorylated to a high basal stoichiometry in striatum, suggestive of basal inhibition of PP2A in striatal neurons. In support of this hypothesis, conditional knock-out of ARPP-16 in CaMKIIα::cre/floxed ARPP-16/19 mice results in dephosphorylation of a subset of PP2A substrates including phospho-Thr75-DARPP-32, phospho-T308-Akt, and phospho-T202/Y204-ERK. Conditional knock-out of ARPP-16/19 is associated with increased motivation measured on a progressive ratio schedule of food reinforcement, yet an attenuated locomotor response to acute cocaine. Our previous studies have shown that ARPP-16 is phosphorylated at Ser88 by PKA. Activation of PKA in striatal slices leads to phosphorylation of Ser88, and this is accompanied by marked dephosphorylation of Ser46. Together, these studies suggest that phospho-Ser46-ARPP-16 acts to basally control PP2A in striatal medium spiny neurons but that dopamine acting via PKA inactivates ARPP-16 leading to selective potentiation of PP2A signaling. SIGNIFICANCE STATEMENT We describe a novel mechanism of signal transduction enriched in medium spiny neurons of striatum that likely mediates effects of the neurotransmitter dopamine acting on these cells. We find that the protein ARPP-16, which is highly expressed in striatal medium spiny neurons, acts as a selective inhibitor of certain forms of the serine/threonine protein phosphatase, PP2A, when phosphorylated by the kinase, MAST3. Under basal conditions, ARPP-16 is phosphorylated by MAST3 to a very high stoichiometry. However, the actions of MAST3 are antagonized by dopamine and cAMP-regulated signaling leading to disinhibition of ARPP-16 and increased PP2A action. PMID:28167675

ARPP-16 Is a Striatal-Enriched Inhibitor of Protein Phosphatase 2A Regulated by Microtubule-Associated Serine/Threonine Kinase 3 (Mast 3 Kinase).

PubMed

Andrade, Erika C; Musante, Veronica; Horiuchi, Atsuko; Matsuzaki, Hideo; Brody, A Harrison; Wu, Terence; Greengard, Paul; Taylor, Jane R; Nairn, Angus C

2017-03-08

ARPP-16 (cAMP-regulated phospho-protein of molecular weight 16 kDa) is one of several small acid-soluble proteins highly expressed in medium spiny neurons of striatum that are phosphorylated in response to dopamine acting via D1 receptor/protein kinase A (PKA) signaling. We show here that ARPP-16 is also phosphorylated in vitro and in vivo by microtubule-associated serine/threonine kinase 3 (MAST3 kinase), an enzyme of previously unknown function that is enriched in striatum. We find that ARPP-16 interacts directly with the scaffolding A subunit of the serine/threonine protein phosphatase, PP2A, and that phosphorylation of ARPP-16 at Ser46 by MAST3 kinase converts the protein into a selective inhibitor of B55α- and B56δ-containing heterotrimeric forms of PP2A. Ser46 of ARPP-16 is phosphorylated to a high basal stoichiometry in striatum, suggestive of basal inhibition of PP2A in striatal neurons. In support of this hypothesis, conditional knock-out of ARPP-16 in CaMKIIα::cre/floxed ARPP-16/19 mice results in dephosphorylation of a subset of PP2A substrates including phospho-Thr75-DARPP-32, phospho-T308-Akt, and phospho-T202/Y204-ERK. Conditional knock-out of ARPP-16/19 is associated with increased motivation measured on a progressive ratio schedule of food reinforcement, yet an attenuated locomotor response to acute cocaine. Our previous studies have shown that ARPP-16 is phosphorylated at Ser88 by PKA. Activation of PKA in striatal slices leads to phosphorylation of Ser88, and this is accompanied by marked dephosphorylation of Ser46. Together, these studies suggest that phospho-Ser46-ARPP-16 acts to basally control PP2A in striatal medium spiny neurons but that dopamine acting via PKA inactivates ARPP-16 leading to selective potentiation of PP2A signaling. SIGNIFICANCE STATEMENT We describe a novel mechanism of signal transduction enriched in medium spiny neurons of striatum that likely mediates effects of the neurotransmitter dopamine acting on these cells. We find that the protein ARPP-16, which is highly expressed in striatal medium spiny neurons, acts as a selective inhibitor of certain forms of the serine/threonine protein phosphatase, PP2A, when phosphorylated by the kinase, MAST3. Under basal conditions, ARPP-16 is phosphorylated by MAST3 to a very high stoichiometry. However, the actions of MAST3 are antagonized by dopamine and cAMP-regulated signaling leading to disinhibition of ARPP-16 and increased PP2A action. Copyright © 2017 the authors 0270-6474/17/372709-14$15.00/0.

Flight Investigation of the Knock-Limited Performance of a Triptane Blend, a Toluene Blend, and 28-R Fuel in an R-1830-75 Engine

NASA Technical Reports Server (NTRS)

Blackman, Calvin C.

1946-01-01

Knock-limited performance data were obtained for three fuels on an R-1830-75 engine in a B-24D airplane at engine speeds of 1800, 2250, and 2600 rpm, a spark advance of 25 degrees B.T.C., and carburetor-air temperatures of 85 F for 1800 and 2250 rpm and 100 F for 2600 rpm. The test fuels were a blend of 80 percent 28-R plus 20 percent triptane (leaded to 4.5 ml TEL/gal), a blend of 80 percent 28-R plus 15 percent toluene (leaded to 4.5 ml TEL / gal), and 28-R fuel. The knock-limited manifold pressure of the toluene blend depreciated more in the lean region than the triptane blend or 28-R fuel. The knock-limited brake horsepower for the triptane blend varied from 16 to 25 percent higher than 28-R in the lean region and 18 to 30 percent higher in the rich region. The knock-limited brake horsepower of the toluene blend was approximately 15 percent higher than that of 28-R in the rich region and varied from 2 to 10 percent higher in the lean region. Knock limits of the triptane blend and 28-R fuel tested in the R-1830-75 engine agreed with limits for the same fuels determined with the R-1830-94 engine for engine speeds of 1800 and 2250 rpm.

CRISPR/Cas9-mediated knock-in of an optimized TetO repeat for live cell imaging of endogenous loci.

PubMed

Tasan, Ipek; Sustackova, Gabriela; Zhang, Liguo; Kim, Jiah; Sivaguru, Mayandi; HamediRad, Mohammad; Wang, Yuchuan; Genova, Justin; Ma, Jian; Belmont, Andrew S; Zhao, Huimin

2018-06-15

Nuclear organization has an important role in determining genome function; however, it is not clear how spatiotemporal organization of the genome relates to functionality. To elucidate this relationship, a method for tracking any locus of interest is desirable. Recently clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) or transcription activator-like effectors were adapted for imaging endogenous loci; however, they are mostly limited to visualization of repetitive regions. Here, we report an efficient and scalable method named SHACKTeR (Short Homology and CRISPR/Cas9-mediated Knock-in of a TetO Repeat) for live cell imaging of specific chromosomal regions without the need for a pre-existing repetitive sequence. SHACKTeR requires only two modifications to the genome: CRISPR/Cas9-mediated knock-in of an optimized TetO repeat and its visualization by TetR-EGFP expression. Our simplified knock-in protocol, utilizing short homology arms integrated by polymerase chain reaction, was successful at labeling 10 different loci in HCT116 cells. We also showed the feasibility of knock-in into lamina-associated, heterochromatin regions, demonstrating that these regions prefer non-homologous end joining for knock-in. Using SHACKTeR, we were able to observe DNA replication at a specific locus by long-term live cell imaging. We anticipate the general applicability and scalability of our method will enhance causative analyses between gene function and compartmentalization in a high-throughput manner.

Characterization of a knock-in mouse model of the homozygous p.V37I variant in Gjb2.

PubMed

Chen, Ying; Hu, Lingxiang; Wang, Xueling; Sun, Changling; Lin, Xin; Li, Lei; Mei, Ling; Huang, Zhiwu; Yang, Tao; Wu, Hao

2016-09-13

The homozygous p.V37I variant in GJB2 is prevalent in East and Southeast Asians and may lead to mild-to-moderate hearing loss with reduced penetrance. To investigate the pathogenic mechanism underlying this variant, we generated a knock-in mouse model of homozygous p.V37I by an embryonic stem cell gene targeting method. Auditory brainstem response test showed that the knock-in mice developed progressive, mild-to-moderate hearing loss over the first 4-9 months. Overall no significant developmental and morphological abnormality was observed in the knock-in mouse cochlea, while confocal immunostaining and electron microscopic scanning revealed minor loss of the outer hair cells. Gene expression microarray analysis identified 105 up-regulated and 43 down-regulated genes in P5 knock-in mouse cochleae (P < 0.05 adjusted by the Benjamini &Hochberg method), among which four top candidate genes with the highest fold-changes or implication to deafness Fcer1g, Nnmt and Lars2 and Cuedc1 were verified by quantitative real-time PCR. Our study demonstrated that the homozygous p.V37I knock-in mouse modeled the hearing phenotype of the human patients and can serve as a useful animal model for further studies. The differentially expressed genes identified in this study may shed new insights into the understanding of the pathogenic mechanism and the phenotypic modification of homozygous p.V37I.

Disruption of KEX1 gene reduces the proteolytic degradation of secreted two-chain Insulin glargine in Pichia pastoris.

PubMed

Sreenivas, Suma; Krishnaiah, Sateesh M; Shyam Mohan, Anil H; Mallikarjun, Niveditha; Govindappa, Nagaraja; Chatterjee, Amarnath; Sastry, Kedarnath N

2016-02-01

Insulin glargine is a slow acting analog of insulin used in diabetes therapy. It is produced by recombinant DNA technology in different hosts namely E. coli and Pichia pastoris. In our previous study, we have described the secretion of fully folded two-chain Insulin glargine into the medium by over-expression of Kex2 protease. The enhanced levels of the Kex2 protease was responsible for the processing of the glargine precursor with in the host. Apart from the two-chain glargine product we observed a small proportion of arginine clipped species. This might be due to the clipping of arginine present at the C-terminus of the B-chain as it is exposed upon Kex2 cleavage. The carboxypeptidase precursor Kex1 is known to be responsible for clipping of C-terminal lysine or arginine of the proteins or peptides. In order to address this issue we created a Kex1 knock out in the host using Cre/loxP mechanism of targeted gene deletion. When two-chain glargine was expressed in the Kex1 knock out host of P. pastoris GS115 the C-terminal clipped species reduced by ∼80%. This modification further improved the process by reducing the levels of product related impurities. Copyright © 2015 Elsevier Inc. All rights reserved.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Parl, Angelika; Mitchell, Sabrina L.; Clay, Hayley B.

Highlights: •The function of the mitochondria fatty acid synthesis pathway is partially unknown. •Overexpression of the pathway causes transcriptional activation through PPARs. •Knock down of the pathway attenuates that activation. •The last enzyme in the pathway regulates its own transcription. •Products of the mtFASII pathway are able to drive nuclear transcription. -- Abstract: Mammalian cells contain two fatty acid synthesis pathways, the cytosolic FASI pathway, and the mitochondrial FASII pathway. The selection behind the conservation of the mitochondrial pathway is not completely understood, given the presence of the cytosolic FAS pathway. In this study, we show through heterologous gene reportermore » systems and PCR-based arrays that overexpression of MECR, the last step in the mtFASII pathway, causes modulation of gene expression through the PPAR pathway. Electromobility shift assays (EMSAs) demonstrate that overexpression of MECR causes increased binding of PPARs to DNA, while cell fractionation and imaging studies show that MECR remains localized to the mitochondria. Interestingly, knock down of the mtFASII pathway lessens the effect of MECR on this transcriptional modulation. Our data are most consistent with MECR-mediated transcriptional activation through products of the mtFASII pathway, although we cannot rule out MECR acting as a coactivator. Further investigation into the physiological relevance of this communication will be necessary to better understand some of the phenotypic consequences of deficits in this pathway observed in animal models and human disease.« less

Pou4f2-GFP knock-in mouse line: A model for studying retinal ganglion cell development.

PubMed

Zheng, Dongwang; Yang, Xiaoyan; Sheng, Donglai; Yu, Dongliang; Liang, Guoqing; Guo, Luming; Xu, Mei; Hu, Xu; He, Daqiang; Yang, Yang; Wang, Yuying

2016-10-01

Pou4f2 acts as a key node in the comprehensive and step-wise gene regulatory network (GRN) and regulates the development of retinal ganglion cells (RGCs). Accordingly, deletion of Pou4f2 results in RGC axon defects and apoptosis. To investigate the GRN involved in RGC regeneration, we generated a mouse line with a POU4F2-green fluorescent protein (GFP) fusion protein expressed in RGCs. Co-localization of POU4F2 and GFP in the retina and brain of Pou4f2-GFP/+ heterozygote mice was confirmed using immunofluorescence analysis. Compared with those in wild-type mice, the expression patterns of POU4F2 and POU4F1 and the co-expression patterns of ISL1 and POU4F2 were unaffected in Pou4f2-GFP/GFP homozygote mice. Moreover, the quantification of RGCs showed no significant difference between Pou4f2-GFP/GFP homozygote and wild-type mice. These results demonstrated that the development of RGCs in Pou4f2-GFP/GFP homozygote mice was the same as in wild-type mice. Thus, the present Pou4f2-GFP knock-in mouse line is a useful tool for further studies on the differentiation and regeneration of RGCs. © 2016 Wiley Periodicals, Inc.

Solar Opportunity and Local Access Rights Act

THOMAS, 112th Congress

Rep. Cardoza, Dennis A. [D-CA-18

2011-04-15

House - 05/02/2011 Referred to the Subcommittee on Insurance, Housing and Community Opportunity. (All Actions) Tracker: This bill has the status IntroducedHere are the steps for Status of Legislation:

76 FR 50269 - Notice of Determinations Regarding Eligibility To Apply for Worker Adjustment Assistance and...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-08-12

... the Andean Trade Preference Act, African Growth and Opportunity Act, or the Caribbean Basin Economic... Trade Act have been met. TA-W-80,137; Yorktowne, Inc., Red Lion, Pennsylvania: March 31, 2010. TA-W-80...

Feedback Activation of Basic Fibroblast Growth Factor Signaling via the Wnt/β-Catenin Pathway in Skin Fibroblasts

PubMed Central

Wang, Xu; Zhu, Yuting; Sun, Congcong; Wang, Tao; Shen, Yingjie; Cai, Wanhui; Sun, Jia; Chi, Lisha; Wang, Haijun; Song, Na; Niu, Chao; Shen, Jiayi; Cong, Weitao; Zhu, Zhongxin; Xuan, Yuanhu; Li, Xiaokun; Jin, Litai

2017-01-01

Skin wound healing is a complex process requiring the coordinated behavior of many cell types, especially in the proliferation and migration of fibroblasts. Basic fibroblast growth factor (bFGF) is a member of the FGF family that promotes fibroblast migration, but the underlying molecular mechanism remains elusive. The present RNA sequencing study showed that the expression levels of several canonical Wnt pathway genes, including Wnt2b, Wnt3, Wnt11, T-cell factor 7 (TCF7), and Frizzled 8 (FZD8) were modified by bFGF stimulation in fibroblasts. Enzyme-linked immunosorbent assay (ELISA) analysis also showed that Wnt pathway was activated under bFGF treatment. Furthermore, treatment of fibroblasts with lithium chloride or IWR-1, an inducer and inhibitor of the Wnt signaling pathway, respectively, promoted and inhibited cell migration. Also, levels of cytosolic glycogen synthase kinase 3 beta phosphorylated at serine9 (pGSK3β Ser9) and nuclear β-catenin were increased upon exposure to bFGF. Molecular and biochemical assays indicated that phosphoinositide 3-kinase (PI3K) signaling activated the GSK3β/β-catenin/Wnt signaling pathway via activation of c-Jun N-terminal kinase (JNK), suggesting that PI3K and JNK act at the upstream of β-catenin. In contrast, knock-down of β-catenin delayed fibroblast cell migration even under bFGF stimulation. RNA sequencing analysis of β-catenin knock-down fibroblasts demonstrated that β-catenin positively regulated the transcription of bFGF and FGF21. Moreover, FGF21 treatment activated AKT and JNK, and accelerated fibroblast migration to a similar extent as bFGF does. In addition, ELISA analysis demonstrated that both of bFGF and FGF21 were auto secretion factor and be regulated by Wnt pathway stimulators. Taken together, our analyses define a feedback regulatory loop between bFGF (FGF21) and Wnt signaling acting through β-catenin in skin fibroblasts. PMID:28217097

Development of a one-step gene knock-out and knock-in method for metabolic engineering of Aureobasidium pullulans.

PubMed

Guo, Jian; Wang, Yuanhua; Li, Baozhong; Huang, Siyao; Chen, Yefu; Guo, Xuewu; Xiao, Dongguang

2017-06-10

Aureobasidium pullulans is an increasingly attractive host for bio-production of pullulan, heavy oil, polymalic acid, and a large spectrum of extracellular enzymes. To date, genetic manipulation of A. pullulans mainly relies on time-consuming conventional restriction enzyme digestion and ligation methods. In this study, we present a one-step homologous recombination-based method for rapid genetic manipulation in A. pullulans. Overlaps measuring >40bp length and 10μg DNA segments for homologous recombination provided maximum benefits to transformation of A. pullulans. This optimized method was successfully applied to PKSIII gene (encodes polyketide synthase) knock-out and gltP gene (encodes glycolipid transfer protein) knock-in. After disruption of PKSIII gene, secretion of melanin decreased slightly. The melanin purified from disruptant showed lower reducing capacity compared with that of the parent strain, leading to a decrease in exopolysaccharide production. Knock-in of gltP gene resulted in at least 4.68-fold increase in heavy oil production depending on the carbon source used, indicating that gltP can regulate heavy oil synthesis in A. pullulans. Copyright © 2017 Elsevier B.V. All rights reserved.

Recapitulating X-Linked Juvenile Retinoschisis in Mouse Model by Knock-In Patient-Specific Novel Mutation.

PubMed

Chen, Ding; Xu, Tao; Tu, Mengjun; Xu, Jinlin; Zhou, Chenchen; Cheng, Lulu; Yang, Ruqing; Yang, Tanchu; Zheng, Weiwei; He, Xiubin; Deng, Ruzhi; Ge, Xianglian; Li, Jin; Song, Zongming; Zhao, Junzhao; Gu, Feng

2017-01-01

X-linked juvenile retinoschisis (XLRS) is a retinal disease caused by mutations in the gene encoding retinoschisin (RS1), which leads to a significant proportion of visual impairment and blindness. To develop personalized genome editing based gene therapy, knock-in animal disease models that have the exact mutation identified in the patients is extremely crucial, and that the way which genome editing in knock-in animals could be easily transferred to the patients. Here we recruited a family diagnosed with XLRS and identified the causative mutation ( RS1 , p.Y65X), then a knock-in mouse model harboring this disease-causative mutation was generated via TALEN (transcription activator-like effector nucleases). We found that the b-wave amplitude of the ERG of the RS1 -KI mice was significantly decreased. Moreover, we observed that the structure of retina in RS1 -KI mice has become disordered, including the disarray of inner nuclear layer and outer nuclear layer, chaos of outer plexiform layer, decreased inner segments of photoreceptor and the loss of outer segments. The novel knock-in mice ( RS1 -KI) harboring patient-specific mutation will be valuable for development of treatment via genome editing mediated gene correction.

Preservation Enhancement and Savings Opportunity Act of 2012

THOMAS, 112th Congress

Rep. Paulsen, Erik [R-MN-3

2012-03-22

House - 04/26/2012 Referred to the Subcommittee on Insurance, Housing and Community Opportunity. (All Actions) Tracker: This bill has the status IntroducedHere are the steps for Status of Legislation:

78 FR 32237 - Draft Initial Comprehensive Plan and Draft Programmatic Environmental Assessment

Federal Register 2010, 2011, 2012, 2013, 2014

2013-05-29

... Resources and Ecosystems Sustainability, Tourist Opportunities, and Revived Economies of the Gulf States Act..., Congress passed and the President signed the Resources and Ecosystems Sustainability, Tourist Opportunities...

40 CFR 91.6 - Reference materials.

Code of Federal Regulations, 2010 CFR

2010-07-01

... Appendix A to Subpart D. ASTM D2699-92: Standard Test Method for Knock Characteristics of Motor Fuels by the Research Method Appendix A to Subpart D. ASTM D2700-92: Standard Test Method for Knock...

29 CFR 1926.350 - Gas welding and cutting.

Code of Federal Regulations, 2011 CFR

2011-07-01

... from being knocked over while in use. (8) When work is finished, when cylinders are empty, or when..., or gangways. Assigned storage places shall be located where cylinders will not be knocked over or...

40 CFR 91.6 - Reference materials.

Code of Federal Regulations, 2011 CFR

2011-07-01

... Appendix A to Subpart D. ASTM D2699-92: Standard Test Method for Knock Characteristics of Motor Fuels by the Research Method Appendix A to Subpart D. ASTM D2700-92: Standard Test Method for Knock...

40 CFR 90.7 - Reference materials.

Code of Federal Regulations, 2011 CFR

2011-07-01

... Appendix A to subpart D, Table 3. ASTM D2699-92: Standard Test Method for Knock Characteristics of Motor... Knock Characteristics of Motor and Aviation Fuels by the Motor Method Appendix A to subpart D, Table 3...

40 CFR 90.7 - Reference materials.

Code of Federal Regulations, 2010 CFR

2010-07-01

... Appendix A to subpart D, Table 3. ASTM D2699-92: Standard Test Method for Knock Characteristics of Motor... Knock Characteristics of Motor and Aviation Fuels by the Motor Method Appendix A to subpart D, Table 3...

Do Structural Missense Variants in the ATM Gene Found in Women With Breast Cancer Cause Breast Cancer in Knock-in Mouse Strains?

DTIC Science & Technology

2006-04-01

W81XWH-05-1-0282 TITLE: Do Structural Missense Variants in the ATM Gene Found in Women with Breast Cancer Cause Breast Cancer in "Knock-in...5a. CONTRACT NUMBER Do Structural Missense Variants in the ATM Gene Found in Women with Breast Cancer Cause Breast Cancer in "Knock-in" Mouse...human cohort-specific missense mutations will develop breast cancer with dominant inheritance in a subset of animals. It also is hypothesized that

Proton Knock-Out in Hall A

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kees de Jager

Proton knock-out is studied in a broad program in Hall A at Jefferson Lab. The first experiment performed in Hall A studied the {sup 16}O(e,e'p) reaction. Since then proton knock-out experiments have studied a variety of aspects of that reaction, from single-nucleon properties to its mechanism, such as final-state interactions and two-body currents, in nuclei from {sup 2}H to {sup 16}O. In this review the results of this program will be summarized and an outlook given of future accomplishments.

Knocking in the Otto-cycle Engine

NASA Technical Reports Server (NTRS)

Weinhart, H

1939-01-01

Engine knock is, as is known, preceded by normal burning of the first part of the charge, and only the part burned last (residual charge), knocks. The aim of the present measurements was, first, to reexamine the combustion form in this residual charge, because of the absence of uniform and frequently contradictory results in the very extensive literature on the subject. On top of that, an attempt was to be made to gain a deeper insight into the mechanism accompanying the combustion process, by means of the electrical test equipment perfected in recent years.

28 CFR 42.702 - Definitions.

Code of Federal Regulations, 2014 CFR

2014-07-01

... Judicial Administration DEPARTMENT OF JUSTICE NONDISCRIMINATION; EQUAL EMPLOYMENT OPPORTUNITY; POLICIES AND... of the Age Discrimination Act of 1975 General Provisions § 42.702 Definitions. As used in this subpart, the term: Act means the Age Discrimination Act of 1975, as amended, 42 U.S.C. 6101-6107. Action...

Expression of inactive glutathione peroxidase 4 leads to embryonic lethality, and inactivation of the Alox15 gene does not rescue such knock-in mice.

PubMed

Brütsch, Simone Hanna; Wang, Chi Chiu; Li, Lu; Stender, Hannelore; Neziroglu, Nilgün; Richter, Constanze; Kuhn, Hartmut; Borchert, Astrid

2015-02-01

Glutathione peroxidases (Gpx) and lipoxygenases (Alox) are functional counterplayers in the metabolism of hydroperoxy lipids that regulate cellular redox homeostasis. Gpx4 is a moonlighting protein that has been implicated not only as an enzyme in anti-oxidative defense, gene expression regulation, and programmed cell death, but also as a structural protein in spermatogenesis. Homozygous Gpx4 knock-out mice are not viable, but molecular reasons for intrauterine lethality are not completely understood. This study was aimed at investigating whether the lack of catalytic activity or the impaired function as structural protein is the dominant reason for embryonic lethality. We further explored whether the pro-oxidative enzyme mouse 12/15 lipoxygenase (Alox15) plays a major role in embryonic lethality of Gpx4-deficient mice. To achieve these goals, we first created knock-in mice, which express a catalytically inactive Gpx4 mutant (Sec46Ala). As homozygous Gpx4-knock-out mice Sec46Ala-Gpx4(+/+) knock-in animals are not viable but undergo intrauterine resorption between embryonic day 6 and 7 (E6-7). In contrast, heterozygous knock-in mice (Sec46Ala-Gpx4(-/+)) are viable, fertile and do not show major phenotypic alterations. Interestingly, homozygous Alox15 deficiency did not rescue the U46A-Gpx4(+/+) mice from embryonic lethality. In fact, when heterozygous U46A-Gpx4(-/+) mice were stepwise crossed into an Alox15-deficent background, no viable U46A-Gpx4(+/+)+Alox15(-/-) individuals were obtained. However, we were able to identify U46A-Gpx4(+/+)+Alox15(-/-) embryos in the state of resorption around E7. These data suggest that the lack of catalytic activity is the major reason for the embryonic lethality of Gpx4(-/-) mice and that systemic inactivation of the Alox15 gene does not rescue homozygous knock-in mice expressing catalytically silent Gpx4.

Synaptic function is modulated by LRRK2 and glutamate release is increased in cortical neurons of G2019S LRRK2 knock-in mice.

PubMed

Beccano-Kelly, Dayne A; Kuhlmann, Naila; Tatarnikov, Igor; Volta, Mattia; Munsie, Lise N; Chou, Patrick; Cao, Li-Ping; Han, Heather; Tapia, Lucia; Farrer, Matthew J; Milnerwood, Austen J

2014-01-01

Mutations in Leucine-Rich Repeat Kinase-2 (LRRK2) result in familial Parkinson's disease and the G2019S mutation alone accounts for up to 30% in some ethnicities. Despite this, the function of LRRK2 is largely undetermined although evidence suggests roles in phosphorylation, protein interactions, autophagy and endocytosis. Emerging reports link loss of LRRK2 to altered synaptic transmission, but the effects of the G2019S mutation upon synaptic release in mammalian neurons are unknown. To assess wild type and mutant LRRK2 in established neuronal networks, we conducted immunocytochemical, electrophysiological and biochemical characterization of >3 week old cortical cultures of LRRK2 knock-out, wild-type overexpressing and G2019S knock-in mice. Synaptic release and synapse numbers were grossly normal in LRRK2 knock-out cells, but discretely reduced glutamatergic activity and reduced synaptic protein levels were observed. Conversely, synapse density was modestly but significantly increased in wild-type LRRK2 overexpressing cultures although event frequency was not. In knock-in cultures, glutamate release was markedly elevated, in the absence of any change to synapse density, indicating that physiological levels of G2019S LRRK2 elevate probability of release. Several pre-synaptic regulatory proteins shown by others to interact with LRRK2 were expressed at normal levels in knock-in cultures; however, synapsin 1 phosphorylation was significantly reduced. Thus, perturbations to the pre-synaptic release machinery and elevated synaptic transmission are early neuronal effects of LRRK2 G2019S. Furthermore, the comparison of knock-in and overexpressing cultures suggests that one copy of the G2019S mutation has a more pronounced effect than an ~3-fold increase in LRRK2 protein. Mutant-induced increases in transmission may convey additional stressors to neuronal physiology that may eventually contribute to the pathogenesis of Parkinson's disease.

Elevated progranulin contributes to synaptic and learning deficit due to loss of fragile X mental retardation protein.

PubMed

Zhang, Kun; Li, Yu-Jiao; Guo, Yanyan; Zheng, Kai-Yin; Yang, Qi; Yang, Le; Wang, Xin-Shang; Song, Qian; Chen, Tao; Zhuo, Min; Zhao, Ming-Gao

2017-12-01

Fragile X syndrome is an inheritable form of intellectual disability caused by loss of fragile X mental retardation protein (FMRP, encoded by the FMR1 gene). Absence of FMRP caused overexpression of progranulin (PGRN, encoded by GRN), a putative tumour necrosis factor receptor ligand. In the present study, we found that progranulin mRNA and protein were upregulated in the medial prefrontal cortex of Fmr1 knock-out mice. In Fmr1 knock-out mice, elevated progranulin caused insufficient dendritic spine pruning and late-phase long-term potentiation in the medial prefrontal cortex of Fmr1 knock-out mice. Partial progranulin knock-down restored spine morphology and reversed behavioural deficits, including impaired fear memory, hyperactivity, and motor inflexibility in Fmr1 knock-out mice. Progranulin increased levels of phosphorylated glutamate ionotropic receptor GluA1 and nuclear factor kappa B in cultured wild-type neurons. Tumour necrosis factor receptor 2 antibody perfusion blocked the effects of progranulin on GluA1 phosphorylation; this result indicates that tumour necrosis factor receptor 2 is required for progranulin-mediated GluA1 phosphorylation and late-phase long-term potentiation expression. However, high basal level of progranulin in Fmr1 knock-out mice prevented further facilitation of synaptic plasticity by exogenous progranulin. Partial downregulation of progranulin or tumour necrosis factor receptor 2/nuclear factor kappa B signalling restored synaptic plasticity and memory deficits in Fmr1 knock-out mice. These findings suggest that elevated PGRN is linked to cognitive deficits of fragile X syndrome, and the progranulin/tumour necrosis factor receptor 2 signalling pathway may be a putative therapeutic target for improving cognitive deficits in fragile X syndrome. © The Author (2017). Published by Oxford University Press on behalf of the Guarantors of Brain. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

40 CFR 35.935-6 - Equal employment opportunity.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 1 2010-07-01 2010-07-01 false Equal employment opportunity. 35.935-6 Section 35.935-6 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GRANTS AND OTHER FEDERAL ASSISTANCE STATE AND LOCAL ASSISTANCE Grants for Construction of Treatment Works-Clean Water Act § 35.935-6 Equal employment opportunity. Contracts...

76 FR 67017 - Praesidian Capital Opportunity Fund III, LP License No. 02/02-0647; Notice Seeking Exemption...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-10-28

... II, LP, Associate of Praesidian Capital Opportunity Fund III, LP, holds a debt investment and warrant... SMALL BUSINESS ADMINISTRATION Praesidian Capital Opportunity Fund III, LP License No. 02/02- 0647; Notice Seeking Exemption Under Section 312 of the Small Business Investment Act, Conflicts of Interest...

Welfare Reform and Postsecondary Education: Research and Policy Update. TANF Requirements Limit Welfare Recipients' Educational Opportunities.

ERIC Educational Resources Information Center

Finney, Johanna

1998-01-01

The Temporary Assistance for Needy Families (TANF) Program work requirements, part of the 1996 Personal Responsibility and Work Opportunity Reconciliation Act (PRWORA) drastically limit women's opportunities to participate in postsecondary education programs while receiving TANF funds. Unlike previous laws governing Aid to Families with Dependent…

29 CFR 452.94 - Reasonable opportunity to vote.

Code of Federal Regulations, 2010 CFR

2010-07-01

... DISCLOSURE ACT OF 1959 Right To Vote § 452.94 Reasonable opportunity to vote. The statutory protection of the right to vote implies that there must be a reasonable opportunity to vote. Thus, there is an obligation... ways, depending on factors such as the distance between the members' work site or homes and the polling...

29 CFR 1614.408 - Civil action: Equal Pay Act.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 29 Labor 4 2010-07-01 2010-07-01 false Civil action: Equal Pay Act. 1614.408 Section 1614.408... EQUAL EMPLOYMENT OPPORTUNITY Appeals and Civil Actions § 1614.408 Civil action: Equal Pay Act. A..., three years of the date of the alleged violation of the Equal Pay Act regardless of whether he or she...

Awareness among Family Members Having Children with Mental Retardation on Relevant Legislations in India

ERIC Educational Resources Information Center

Kuppusamy, Balabaskar; Narayan, Jayanthi; Nair, Deepa

2012-01-01

In India, Acts and legislations are in place for persons with mental retardation. Three major Acts namely Rehabilitation Council of India Act (1992), Persons with Disabilities (Equal opportunities, protection of rights and full participation) Act (1995) and National Trust for welfare of persons with Autism, Cerebral Palsy, Mental retardation and…

The Great Balancing Act: Financial Equity and Local Control. Can Vermont's Act 60 Do It?

ERIC Educational Resources Information Center

Jimerson, Lorna

Vermont's Equal Educational Opportunity Act of 1997 (Act 60) has attracted attention because of its funding and quality components, and the highly publicized responses to its implementation. A key feature is its "recapture" provision in which property-wealthy towns contribute to a state educational fund that helps support education in…

Opportunity and Danger: The Two Sides of the American Recovery and Reinvestment Act

ERIC Educational Resources Information Center

Warner, Carolyn

2009-01-01

The American Recovery and Reinvestment Act (ARRA) is likely the greatest single infusion of federal dollars into education since Congress passed the Elementary and Secondary Education Act in 1965 during Lyndon Johnson's presidency. When President Johnson signed that act, he said its purpose was to give every child in America "all the…

Knock-Outs, Stick-Outs, Cut-Outs: Clipping Paths Separate Objects from Background.

ERIC Educational Resources Information Center

Wilson, Bradley

1998-01-01

Outlines a six-step process that allows computer operators, using Photoshop software, to create "knock-outs" to precisely define the path that will serve to separate the object from the background. (SR)

Long noncoding RNA lnc-sox5 modulates CRC tumorigenesis by unbalancing tumor microenvironment.

PubMed

Wu, Kaiming; Zhao, Zhenxian; Liu, Kuanzhi; Zhang, Jian; Li, Guanghua; Wang, Liang

2017-07-03

Long non-coding RNAs (LncRNAs) have been recently regarded as systemic regulators in multiple biologic processes including tumorigenesis. In this study, we observed the expression of lncRNA lnc-sox5 was significantly increased in colorectal cancer (CRC). Despite the CRC cell growth, cell cycle and cell apoptosis was not affected by lnc-sox5 knock-down, lnc-sox5 knock-down suppressed CRC cell migration and invasion. In addition, xenograft animal model suggested that lnc-sox5 knock-down significantly suppressed the CRC tumorigenesis. Our results also showed that the expression of indoleamine 2,3-dioxygenase 1 (IDO1) was significantly reduced by lnc-sox5 knock-down and therefore modulated the infiltration and cytotoxicity of CD3 + CD8 + T cells. Taken together, these results suggested that lnc-sox5 unbalances tumor microenvironment to regulate colorectal cancer progression.

A High-Speed Motion-Picture Study of Normal Combustion, Knock and Preignition in a Spark-Ignition Engines

NASA Technical Reports Server (NTRS)

Rothrock, A M; Spencer, R C; Miller, Cearcy D

1941-01-01

Combustion in a spark-ignition engine was investigated by means of the NACA high-speed motion-picture cameras. This camera is operated at a speed of 40,000 photographs a second and therefore makes possible the study of changes that take place in the intervals as short as 0.000025 second. When the motion pictures are projected at the normal speed of 16 frames a second, any rate of movement shown is slowed down 2500 times. Photographs are presented of normal combustion, of combustion from preignitions, and of knock both with and without preignition. The photographs of combustion show that knock may be preceded by a period of exothermic reaction in the end zone that persists for a time interval of as much as 0.0006 second. The knock takes place in 0.00005 second or less.

Taking Science to Special Needs and At-Risk Youth in the San Francisco Bay Area

NASA Astrophysics Data System (ADS)

Scherrer, D. K.; Yager, D.; Blair, J.; McCully, D.; Alameda, E.; Crawford, K.

2009-12-01

Youth in Juvenile Detention Facilities do attend (in-house) school, but rarely receive any instruction in science. We report on a new program to bring science to students at the Santa Clara County Juvenile Detention Facilitirs. Working in partnership with the Knock Out Dog Fighting campaign put together by Kris Crawford of For Pits Sake, Inc., our program provides alternatives to inappropriate behavior so often seen in inner city environments by introducing students to hands-on, inquiry based science activities. Likewise, we report on using similar materials to provide hands-on science activities to special needs students in Santa Clara and Santa Cruz Counties through “Take Flight for Kids” events organized by Dean McCully of Cisco Systems. Through “Take Flight for Kids”, amateur pilots offer special needs students rides in light aircraft and invite them to explore science and other activities through a community fair hosted by hundreds of local organizations. The fair highlights science opportunities and is supported and attended by a wide variety of NASA, NSF, and other science-related organizations. Our poster will focus on techniques and materials we use to excite special young people about science and opportunities for them in STEM fields.

Opportunity Knocks: HIV Prevention in Primary Care.

PubMed

Thrun, Mark W

2014-06-01

Expansions in health care coverage, a comprehensive framework for HIV prevention and care, electronic medical records, and novel HIV prevention modalities create a current opportunity to change the trajectory of the HIV epidemic in the United States. HIV is increasingly disproportionately found in populations historically at higher risk, including gay men and other men who have sex with men, transgender women, injection drug users, and persons of color. This underscores the need for providers to identify persons at higher risk for HIV and assure the provision of screening and prevention services. In turn, universal screening for HIV-testing every adolescent and adult at least once in their lifetime-will increasingly be necessary to find the infrequent cases of HIV in lower risk populations. In both these domains, primary care providers will play a unique role in complementing traditional providers of HIV prevention and care services by increasing the proportion of their patients who have been screened for HIV, opening dialogues around sexual health, including asking about sexual orientation and gender identity, and prescribing antivirals as pre- and postexposure prophylaxis for their non-HIV-infected patients. Primary care providers must understand and embrace their importance along the HIV prevention and care continuum.

29 CFR 1620.27 - Relationship to the Equal Pay Act of title VII of the Civil Rights Act.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 29 Labor 4 2010-07-01 2010-07-01 false Relationship to the Equal Pay Act of title VII of the Civil... OPPORTUNITY COMMISSION THE EQUAL PAY ACT § 1620.27 Relationship to the Equal Pay Act of title VII of the Civil... equal pay under the Equal Pay Act has no relationship to whether the employee is in the lower paying job...

Age- and region-specific imbalances of basal amino acids and monoamine metabolism in limbic regions of female Fmr1 knock-out mice.

PubMed

Gruss, Michael; Braun, Katharina

2004-07-01

The Fragile X syndrome, a common form of mental retardation in humans, originates from the loss of expression of the Fragile X mental retardation gene leading to the absence of the encoded Fragile X mental retardation protein 1 (FMRP). A broad pattern of morphological and behavioral abnormalities is well described for affected humans as well as Fmr1 knock-out mice, a transgenic animal model for the human Fragile X syndrome. In the present study, we examined neurochemical differences between female Fmr1 knock-out and wildtype mice with particular focus on neurotransmission. Significant age- and region-specific differences of basal tissue neurotransmitter and metabolite levels measured by high performance liquid chromatography were found. Those differences were more numerous in juvenile animals (postnatal day (PND) 28-31) compared to adults (postnatal day 209-221). In juvenile female knock-out mice, especially aspartate and taurine were increased in cortical regions, striatum, cerebellum, and brainstem. Furthermore, compared to the wildtype animals, the juvenile knock-out mice displayed an increased level of neuronal inhibition in the hippocampus and brainstem reflected by decreased ratios of (aspartate + glutamate)/(taurine + GABA), as well as an increased dopamine (DA) turnover in cortical regions, striatum, and hippocampus. These results provide the first evidence that the lack of FMRP expression in female Fmr1 knock-out mice is accompanied by age-dependent, region-specific alterations in brain amino acids, and monoamine turnover, which might be related to the reported synaptical and behavioural alterations in these animals.

Starch Binding Domain-containing Protein 1 Plays a Dominant Role in Glycogen Transport to Lysosomes in Liver*

PubMed Central

Sun, Tao; Yi, Haiqing; Yang, Chunyu; Kishnani, Priya S.; Sun, Baodong

2016-01-01

A small portion of cellular glycogen is transported to and degraded in lysosomes by acid α-glucosidase (GAA) in mammals, but it is unclear why and how glycogen is transported to the lysosomes. Stbd1 has recently been proposed to participate in glycogen trafficking to lysosomes. However, our previous study demonstrated that knockdown of Stbd1 in GAA knock-out mice did not alter lysosomal glycogen storage in skeletal muscles. To further determine whether Stbd1 participates in glycogen transport to lysosomes, we generated GAA/Stbd1 double knock-out mice. In fasted double knock-out mice, glycogen accumulation in skeletal and cardiac muscles was not affected, but glycogen content in liver was reduced by nearly 73% at 3 months of age and by 60% at 13 months as compared with GAA knock-out mice, indicating that the transport of glycogen to lysosomes was suppressed in liver by the loss of Stbd1. Exogenous expression of human Stbd1 in double knock-out mice restored the liver lysosomal glycogen content to the level of GAA knock-out mice, as did a mutant lacking the Atg8 family interacting motif (AIM) and another mutant that contains only the N-terminal 24 hydrophobic segment and the C-terminal starch binding domain (CBM20) interlinked by an HA tag. Our results demonstrate that Stbd1 plays a dominant role in glycogen transport to lysosomes in liver and that the N-terminal transmembrane region and the C-terminal CBM20 domain are critical for this function. PMID:27358407

29 CFR 1626.1 - Purpose.

Code of Federal Regulations, 2011 CFR

2011-07-01

... to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN... the Equal Employment Opportunity Commission for carrying out its responsibilities in the administration and enforcement of the Age Discrimination in Employment Act of 1967, as amended. ...

29 CFR 1626.1 - Purpose.

Code of Federal Regulations, 2014 CFR

2014-07-01

... to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN... the Equal Employment Opportunity Commission for carrying out its responsibilities in the administration and enforcement of the Age Discrimination in Employment Act of 1967, as amended. ...

29 CFR 1626.1 - Purpose.

Code of Federal Regulations, 2010 CFR

2010-07-01

... to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN... the Equal Employment Opportunity Commission for carrying out its responsibilities in the administration and enforcement of the Age Discrimination in Employment Act of 1967, as amended. ...

29 CFR 1626.1 - Purpose.

Code of Federal Regulations, 2013 CFR

2013-07-01

... to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN... the Equal Employment Opportunity Commission for carrying out its responsibilities in the administration and enforcement of the Age Discrimination in Employment Act of 1967, as amended. ...

29 CFR 1626.1 - Purpose.

Code of Federal Regulations, 2012 CFR

2012-07-01

... to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN... the Equal Employment Opportunity Commission for carrying out its responsibilities in the administration and enforcement of the Age Discrimination in Employment Act of 1967, as amended. ...

76 FR 6469 - Agency Information Collection Activities

Federal Register 2010, 2011, 2012, 2013, 2014

2011-02-04

... EQUAL EMPLOYMENT OPPORTUNITY COMMISSION Agency Information Collection Activities AGENCY: Equal Employment Opportunity Commission. ACTION: Notice of Information Collection--Extension Without Change: Employer Information Report (EEO-1). SUMMARY: In accordance with the Paperwork Reduction Act, the Equal...

40 CFR 745.65 - Lead-based paint hazards.

Code of Federal Regulations, 2010 CFR

2010-07-01

... caused by impact from a related building component (such as a door knob that knocks into a wall or a door that knocks against its door frame. (3) Any chewable lead-based painted surface on which there is...

40 CFR 745.65 - Lead-based paint hazards.

Code of Federal Regulations, 2011 CFR

2011-07-01

... caused by impact from a related building component (such as a door knob that knocks into a wall or a door that knocks against its door frame. (3) Any chewable lead-based painted surface on which there is...

Functional characterization of three MicroRNAs of the Asian Tiger Mosquito, Aedes albopictus

PubMed Central

2013-01-01

Background Temporal and stage specific expression of microRNAs (miRNAs) in embryos, larvae, pupae and adults of Aedes albopictus showed differential expression levels across the four developmental stages, indicating their potential regulatory roles in mosquito development. The functional characterization of these miRNAs was not known. Accordingly our study evaluated the functional characterization of three miRNAs, which are temporally up-regulated in the various developmental stages of Ae. albopictus mosquitoes. Methods miRNA mimics, inhibitors and negative controls were designed and their knock-in and knock-down efficiency were analyzed by qRT-PCR after transfecting the mosquito cell lines C6/36, and also by injecting in their specific developmental stages. The functional role of each individual miRNA was analyzed with various parameters of development such as, hatching rate and hatching time in embryos, eclosion rate in larvae, longevity and fecundity in the adult mosquitoes. Results The knock-in with the specifically designed miRNA mimics showed increased levels of expression of miRNA compared with their normal controls. We confirmed these findings using qRT-PCR, both by in vitro expression in C6/36 mosquito cell lines after transfection as well as in in vivo expression in developmental stages of mosquitoes by microinjection. The knock-down of expression with the corresponding inhibitors showed a considerable decrease in the expression levels of these miRNAs and obvious functional effects in Ae. albopictus development, detected by a decrease in the hatching rate of embryos and eclosion rate in larvae and a marked reduction in longevity and fecundity in adults. Conclusion This study carried out by knock-in and knock-down of specifically and temporally expressed miRNAs in Ae. albopictus by microinjection is a novel study to delineate the importance of the miRNA expression in regulating mosquito development. The knock-down and loss of function of endogenously expressed miRNAs by the miRNA inhibitors in specific developmental stages had considerable effects on development, but enhancement of their gain of function was not observed on knock-in of these specific miRNAs. Hence, our study indicates that an optimal level of endogenous expression of miRNA is indispensable for the normal development and maintenance of the vectorial population density and pathogen transmissibility of this mosquito vector. PMID:23924583

When Opportunity Knocks Twice: Dual Living Kidney Donation, Autonomy and the Public Interest.

PubMed

Bailey, Phillippa; Huxtable, Richard

2016-02-01

Living kidney transplantation offers the best treatment in terms of life-expectancy and quality of life for those with end-stage renal disease. The long-term risks of living donor nephrectomy, although real, are very small, with evidence of good medium-term outcomes. Who should be entitled to donate, and in which circumstances, is nevertheless a live question. We explore the ethical dimensions of a request by an individual to donate both of their kidneys during life: 'dual living kidney donation'. Our ethical analysis is tethered to a hypothetical case study in which a father asks to donate a kidney to each of his twin boys. We explore the autonomy of the protagonists, alongside different dimensions of the public interest, such as the need to protect not only the recipients, but also the donor and even the wider community. Whilst acknowledging objections to 'dual-donation', not least by reference to the harms that the donor might be expected to endure, we suggest there is a prima facie case for permitting this, provided that both donor and recipients are willing and that due attention is paid to such considerations as the autonomy and welfare of all parties, as well as to the wider ramifications of acting on such a request. We argue for broader interpretations of the concepts of autonomy and welfare, recognizing the importance of relationships and the relevance of more than merely physical well-being. Equipped with such a holistic assessment, we suggest there is a prima facie case for allowing 'dual living kidney donation'. © 2015 The Authors. Bioethics Published by John Wiley & Sons Ltd.

Oversight Hearing on the Job Training Partnership Act (Part 1). Hearing before the Subcommittee on Employment Opportunities of the Committee on Education and Labor, House of Representatives, Ninety-Ninth Congress, First Session (Washington, DC, May 2, 1985).

ERIC Educational Resources Information Center

Congress of the U.S., Washington, DC. House Committee on Education and Labor.

This document is a transcript of an oversight hearing on the Job Training Partnership Act (JTPA). It is the first in a series designed by the House Subcommittee on Employment Opportunities, Committee on Education and Labor, to evaluate the program. In his introduction, the subcommittee chairman voiced his concern about proposed budget cuts in…

Opportunities for oncology in the Patient Protection and Affordable Care Act.

PubMed

Patel, Kavita K; Tran, Lisa

2013-01-01

The Patient Protection and Affordable Care Act (ACA) contains within it three significant legislative constructs: to enhance access to health care, improve quality, and decrease cost. Also known as the Triple Aim, these three simple, yet monumental, goals have been the object of actions to date as well as future implementation efforts. This article will identify sections of the legislation that would directly provide areas of opportunity to improve health and achieve the triple aim for the oncology profession.

College Opportunity Act of 1978. Joint Hearing Before the Committee on Human Resources, United States Senate and the Committee on Education and Labor, House of Representatives, Ninety-fifth Congress, Second Session on S. 2539 (February 9, 1978).

ERIC Educational Resources Information Center

Congress of the U.S., Washington, DC. House Committee on Education and Labor.

The transcript of joint hearings on college assistance legislation before the Senate Committee on Human Resources and the House Committee on Education and Labor is presented. The hearings concern amendment of the Higher Education Act of 1965 to improve the Basic Educational Opportunity Grant (BEOG) program. Witnesses included Joseph A. Califano,…

RNA therapeutics targeting osteoclast-mediated excessive bone resorption

PubMed Central

Wang, Yuwei; Grainger, David W

2011-01-01

RNA interference (RNAi) is a sequence-specific post-transcriptional gene silencing technique developed with dramatically increasing utility for both scientific and therapeutic purposes. Short interfering RNA (siRNA) is currently exploited to regulate protein expression relevant to many therapeutic applications, and commonly used as a tool for elucidating disease-associated genes. Osteoporosis and their associated osteoporotic fragility fractures in both men and women are rapidly becoming a global healthcare crisis as average life expectancy increases worldwide. New therapeutics are needed for this increasing patient population. This review describes the diversity of molecular targets suitable for RNAi-based gene knock-down in osteoclasts to control osteoclast-mediated excessive bone resorption. We identify strategies for developing targeted siRNA delivery and efficient gene silencing, and describe opportunities and challenges of introducing siRNA as a therapeutic approach to hard and connective tissue disorders. PMID:21945356

Public Notice 2017-29: Regency Development

EPA Pesticide Factsheets

EPA is providing notice of a proposed administrative penalty assessment for alleged violations of the Clean Water Act (Act). EPA is also providing notice of opportunity to comment on the proposed penalty assessment.

50 CFR 12.25 - Transfers in settlement of civil penalty claims.

Code of Federal Regulations, 2010 CFR

2010-10-01

... may be liable for civil penalty under the Endangered Species Act, 16 U.S.C. 1531 et seq.; Lacey Act, 18 U.S.C. 43; Lacey Act Amendments of 1981, 16 U.S.C. 3371 et seq.; Eagle Protection Act, 16 U.S.C. 668 et seq.; or Marine Mammal Protection Act, 16 U.S.C. 1361 et seq., may be given an opportunity to...

50 CFR 12.25 - Transfers in settlement of civil penalty claims.

Code of Federal Regulations, 2011 CFR

2011-10-01

... may be liable for civil penalty under the Endangered Species Act, 16 U.S.C. 1531 et seq.; Lacey Act, 18 U.S.C. 43; Lacey Act Amendments of 1981, 16 U.S.C. 3371 et seq.; Eagle Protection Act, 16 U.S.C. 668 et seq.; or Marine Mammal Protection Act, 16 U.S.C. 1361 et seq., may be given an opportunity to...

50 CFR 12.25 - Transfers in settlement of civil penalty claims.

Code of Federal Regulations, 2014 CFR

2014-10-01

... may be liable for civil penalty under the Endangered Species Act, 16 U.S.C. 1531 et seq.; Lacey Act, 18 U.S.C. 43; Lacey Act Amendments of 1981, 16 U.S.C. 3371 et seq.; Eagle Protection Act, 16 U.S.C. 668 et seq.; or Marine Mammal Protection Act, 16 U.S.C. 1361 et seq., may be given an opportunity to...

50 CFR 12.25 - Transfers in settlement of civil penalty claims.

Code of Federal Regulations, 2012 CFR

2012-10-01

... may be liable for civil penalty under the Endangered Species Act, 16 U.S.C. 1531 et seq.; Lacey Act, 18 U.S.C. 43; Lacey Act Amendments of 1981, 16 U.S.C. 3371 et seq.; Eagle Protection Act, 16 U.S.C. 668 et seq.; or Marine Mammal Protection Act, 16 U.S.C. 1361 et seq., may be given an opportunity to...

50 CFR 12.25 - Transfers in settlement of civil penalty claims.

Code of Federal Regulations, 2013 CFR

2013-10-01

... may be liable for civil penalty under the Endangered Species Act, 16 U.S.C. 1531 et seq.; Lacey Act, 18 U.S.C. 43; Lacey Act Amendments of 1981, 16 U.S.C. 3371 et seq.; Eagle Protection Act, 16 U.S.C. 668 et seq.; or Marine Mammal Protection Act, 16 U.S.C. 1361 et seq., may be given an opportunity to...

29 CFR 1626.4 - Information concerning alleged violations of the Act.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 29 Labor 4 2014-07-01 2014-07-01 false Information concerning alleged violations of the Act. 1626.4 Section 1626.4 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN EMPLOYMENT ACT § 1626.4 Information concerning alleged violations...

29 CFR 1626.4 - Information concerning alleged violations of the Act.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 29 Labor 4 2010-07-01 2010-07-01 false Information concerning alleged violations of the Act. 1626.4 Section 1626.4 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN EMPLOYMENT ACT § 1626.4 Information concerning alleged violations...

29 CFR 1626.4 - Information concerning alleged violations of the Act.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 29 Labor 4 2013-07-01 2013-07-01 false Information concerning alleged violations of the Act. 1626.4 Section 1626.4 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN EMPLOYMENT ACT § 1626.4 Information concerning alleged violations...

29 CFR 1626.4 - Information concerning alleged violations of the Act.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 29 Labor 4 2011-07-01 2011-07-01 false Information concerning alleged violations of the Act. 1626.4 Section 1626.4 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN EMPLOYMENT ACT § 1626.4 Information concerning alleged violations...

29 CFR 1626.4 - Information concerning alleged violations of the Act.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 29 Labor 4 2012-07-01 2012-07-01 false Information concerning alleged violations of the Act. 1626.4 Section 1626.4 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN EMPLOYMENT ACT § 1626.4 Information concerning alleged violations...

76 FR 20352 - Notice of Intent To Award Affordable Care Act (ACA) Funding

Federal Register 2010, 2011, 2012, 2013, 2014

2011-04-12

... Intent To Award Affordable Care Act (ACA) Funding Notice of Intent to award Affordable Care Act (ACA) funding to two Emerging Infections Program (EIP) grantees, the Connecticut Department of Public Health and... grantees' Fiscal Year (FY) 2011 non-competitive continuation applications under funding opportunity CI05...

76 FR 20354 - Notice of Intent To Award Supplemental Affordable Care Act Funding

Federal Register 2010, 2011, 2012, 2013, 2014

2011-04-12

... Intent To Award Supplemental Affordable Care Act Funding Notice of Intent to award supplemental Affordable Care Act funding to support enhancement of an existing laboratory fellowship training program through funding opportunity CDC-RFA-HM10-1001, ``APHL--CDC Partnership for Quality Laboratory Practice...

29 CFR 1610.17 - Exemptions.

Code of Federal Regulations, 2010 CFR

2010-07-01

... Equal Pay Act (29 U.S.C. 206(b)) or the Age Discrimination in Employment Act of 1967 (29 U.S.C. 621 et... Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION AVAILABILITY OF RECORDS Production or... pursuant to its authority under section 709 prior to the institution of any proceeding under the act...

29 CFR 34.43 - Complaints and investigations.

Code of Federal Regulations, 2010 CFR

2010-07-01

... Equal Pay Act of 1963, as amended (29 U.S.C. 206(d)), or the Age Discrimination in Employment Act of... Office of the Secretary of Labor IMPLEMENTATION OF THE NONDISCRIMINATION AND EQUAL OPPORTUNITY REQUIREMENTS OF THE JOB TRAINING PARTNERSHIP ACT OF 1982, AS AMENDED (JTPA) Compliance Procedures § 34.43...

29 CFR 1620.32 - Recordkeeping requirements.

Code of Federal Regulations, 2010 CFR

2010-07-01

... Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION THE EQUAL PAY ACT § 1620.32 Recordkeeping requirements. (a) Employers having employees subject to the Act are required to keep records in... subject to the equal pay provisions of the Act shall maintain and preserve all records required by the...

29 CFR 1620.34 - Rules to be liberally construed.

Code of Federal Regulations, 2010 CFR

2010-07-01

... Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION THE EQUAL PAY ACT § 1620.34 Rules to be liberally construed. (a) These rules and regulations shall be liberally construed to effectuate the purpose and provisions of this Act and any other Act administered by the...

75 FR 21280 - National Advisory Committee on Institutional Quality and Integrity (NACIQI) Meeting

Federal Register 2010, 2011, 2012, 2013, 2014

2010-04-23

...)(2) of the Federal Advisory Committee Act, section 114(d)(1)(B) of the Higher Education Opportunity Act, and section 114(d)(1)(B) and (D) of the Higher Education Act, as amended. In all instances... the Higher Education Act of 1965, as amended, and 34 CFR parts 602 and 603. In addition, comments for...

Improving the Timeliness of Equal Employment Opportunity Complaint Processing in Department of Defense

DTIC Science & Technology

2015-01-01

1964 (Title VII) and the Pregnancy Discrimination Act amendment to Title VII, the Equal Pay Act of 1963, the Age Discrimi- nation in Employment Act of...Act of 1964 (Title VII) and the Pregnancy Discrimination Act amendment to Title VII, the Equal Pay Act of 1963, the Age Discrimination in...EEO programs uti - lize training on the EEO complaint process and framing of claims and that they use more-structured investigation requests

Hard Religious Questions in "Knee-Knock Rise" and "Tuck Everlasting."

ERIC Educational Resources Information Center

Milner, Joseph O.

1995-01-01

Discusses the way Natalie Babbitt brings up issues of religious conflict and questions about religious certainty and skepticism in her young adult novels. Pays particular attention to two of her novels: "Knee-Knock Rise" and "Tuck Everlasting." (HB)

When Your Back Hurts: Don't Let Back Pain Knock You Flat

MedlinePlus

... Your Back Hurts Don’t Let Back Pain Knock You Flat En español Send us your comments ... Complementary Health Approaches Halt the Hurt! References The SPORT Value Compass: Do the Extra Costs of Undergoing ...

[Synapse maturation and autism: learning from neuroligin model mice].

PubMed

Tabuchi, Katsuhiko; Chang, WenHsin; Hang, WenHsin; Asgar, Nur Farehan; Asgar, Nur Farehan Mohamed; Pramanik, Gopal

2014-02-01

Autism is a neurodevelopmental disorder characterized by impairments in social interaction, communication, and restricted and repetitive behavior. Synaptic defects have been implicated in autism; nevertheless, the cause is still largely unknown. A mutation that substitutes cysteine for arginine at residue 451 of Neuroligin-3 (R451C) is the first monogenic mutation identified in idiopathic autism patients. To study the relationship between this mutation and autism, we generated knock-in mice that recapitulated this mutation. The knock-in mice were born and grew up normally without showing any major physical phenotypes, but showed a deficit in social interaction. We studied synaptic function in the layer II/III pyramidal neurons in the somatosensory cortex and found inhibitory synaptic transmission was enhanced in the knock-in mice. The administration of GABA blocker rescued social interaction, suggesting that this caused autistic behavior in these mice. We also found, by Morris water maze test, that spatial learning and memory were significantly enhanced in the knock-in mice. Electrophysiology in the CA1 region of the hippocampus revealed that LTP, the NMDA/AMPA ratio, and NR2B function were enhanced, indicating that synaptic maturation was impaired in the knock-in mice. This may cause the deficit in social behavior and extraordinary memory ability occasionally seen in autistic patients.

True-breeding targeted gene knock-out in barley using designer TALE-nuclease in haploid cells.

PubMed

Gurushidze, Maia; Hensel, Goetz; Hiekel, Stefan; Schedel, Sindy; Valkov, Vladimir; Kumlehn, Jochen

2014-01-01

Transcription activator-like effector nucleases (TALENs) are customizable fusion proteins able to cleave virtually any genomic DNA sequence of choice, and thereby to generate site-directed genetic modifications in a wide range of cells and organisms. In the present study, we expressed TALENs in pollen-derived, regenerable cells to establish the generation of instantly true-breeding mutant plants. A gfp-specific TALEN pair was expressed via Agrobacterium-mediated transformation in embryogenic pollen of transgenic barley harboring a functional copy of gfp. Thanks to the haploid nature of the target cells, knock-out mutations were readily detected, and homozygous primary mutant plants obtained following genome duplication. In all, 22% of the TALEN transgenics proved knocked out with respect to gfp, and the loss of function could be ascribed to the deletions of between four and 36 nucleotides in length. The altered gfp alleles were transmitted normally through meiosis, and the knock-out phenotype was consistently shown by the offspring of two independent mutants. Thus, here we describe the efficient production of TALEN-mediated gene knock-outs in barley that are instantaneously homozygous and non-chimeric in regard to the site-directed mutations induced. This TALEN approach has broad applicability for both elucidating gene function and tailoring the phenotype of barley and other crop species.

Fluorescent protein tagging of endogenous protein in brain neurons using CRISPR/Cas9-mediated knock-in and in utero electroporation techniques

PubMed Central

Uemura, Takeshi; Mori, Takuma; Kurihara, Taiga; Kawase, Shiori; Koike, Rie; Satoga, Michiru; Cao, Xueshan; Li, Xue; Yanagawa, Toru; Sakurai, Takayuki; Shindo, Takayuki; Tabuchi, Katsuhiko

2016-01-01

Genome editing is a powerful technique for studying gene functions. CRISPR/Cas9-mediated gene knock-in has recently been applied to various cells and organisms. Here, we successfully knocked in an EGFP coding sequence at the site immediately after the first ATG codon of the β-actin gene in neurons in the brain by the combined use of the CRISPR/Cas9 system and in utero electroporation technique, resulting in the expression of the EGFP-tagged β-actin protein in cortical layer 2/3 pyramidal neurons. We detected EGFP fluorescence signals in the soma and neurites of EGFP knock-in neurons. These signals were particularly abundant in the head of dendritic spines, corresponding to the localization of the endogenous β-actin protein. EGFP knock-in neurons showed no detectable changes in spine density and basic electrophysiological properties. In contrast, exogenously overexpressed EGFP-β-actin showed increased spine density and EPSC frequency, and changed resting membrane potential. Thus, our technique provides a potential tool to elucidate the localization of various endogenous proteins in neurons by epitope tagging without altering neuronal and synaptic functions. This technique can be also useful for introducing a specific mutation into genes to study the function of proteins and genomic elements in brain neurons. PMID:27782168

Intracellular generation of single-strand template increases the knock-in efficiency by combining CRISPR/Cas9 with AAV.

PubMed

Xiao, Qing; Min, Taishan; Ma, Shuangping; Hu, Lingna; Chen, Hongyan; Lu, Daru

2018-04-18

Targeted integration of transgenes facilitates functional genomic research and holds prospect for gene therapy. The established microhomology-mediated end-joining (MMEJ)-based strategy leads to the precise gene knock-in with easily constructed donor, yet the limited efficiency remains to be further improved. Here, we show that single-strand DNA (ssDNA) donor contributes to efficient increase of knock-in efficiency and establishes a method to achieve the intracellular linearization of long ssDNA donor. We identified that the CRISPR/Cas9 system is responsible for breaking double-strand DNA (dsDNA) of palindromic structure in inverted terminal repeats (ITRs) region of recombinant adeno-associated virus (AAV), leading to the inhibition of viral second-strand DNA synthesis. Combing Cas9 plasmids targeting genome and ITR with AAV donor delivery, the precise knock-in of gene cassette was achieved, with 13-14% of the donor insertion events being mediated by MMEJ in HEK 293T cells. This study describes a novel method to integrate large single-strand transgene cassettes into the genomes, increasing knock-in efficiency by 13.6-19.5-fold relative to conventional AAV-mediated method. It also provides a comprehensive solution to the challenges of complicated production and difficult delivery with large exogenous fragments.

Fluorescent protein tagging of endogenous protein in brain neurons using CRISPR/Cas9-mediated knock-in and in utero electroporation techniques.

PubMed

Uemura, Takeshi; Mori, Takuma; Kurihara, Taiga; Kawase, Shiori; Koike, Rie; Satoga, Michiru; Cao, Xueshan; Li, Xue; Yanagawa, Toru; Sakurai, Takayuki; Shindo, Takayuki; Tabuchi, Katsuhiko

2016-10-26

Genome editing is a powerful technique for studying gene functions. CRISPR/Cas9-mediated gene knock-in has recently been applied to various cells and organisms. Here, we successfully knocked in an EGFP coding sequence at the site immediately after the first ATG codon of the β-actin gene in neurons in the brain by the combined use of the CRISPR/Cas9 system and in utero electroporation technique, resulting in the expression of the EGFP-tagged β-actin protein in cortical layer 2/3 pyramidal neurons. We detected EGFP fluorescence signals in the soma and neurites of EGFP knock-in neurons. These signals were particularly abundant in the head of dendritic spines, corresponding to the localization of the endogenous β-actin protein. EGFP knock-in neurons showed no detectable changes in spine density and basic electrophysiological properties. In contrast, exogenously overexpressed EGFP-β-actin showed increased spine density and EPSC frequency, and changed resting membrane potential. Thus, our technique provides a potential tool to elucidate the localization of various endogenous proteins in neurons by epitope tagging without altering neuronal and synaptic functions. This technique can be also useful for introducing a specific mutation into genes to study the function of proteins and genomic elements in brain neurons.

Optimization of a CRISPR/Cas9-mediated Knock-in Strategy at the Porcine Rosa26 Locus in Porcine Foetal Fibroblasts.

PubMed

Xie, Zicong; Pang, Daxin; Wang, Kankan; Li, Mengjing; Guo, Nannan; Yuan, Hongming; Li, Jianing; Zou, Xiaodong; Jiao, Huping; Ouyang, Hongsheng; Li, Zhanjun; Tang, Xiaochun

2017-06-08

Genetically modified pigs have important roles in agriculture and biomedicine. However, genome-specific knock-in techniques in pigs are still in their infancy and optimal strategies have not been extensively investigated. In this study, we performed electroporation to introduce a targeting donor vector (a non-linearized vector that did not contain a promoter or selectable marker) into Porcine Foetal Fibroblasts (PFFs) along with a CRISPR/Cas9 vector. After optimization, the efficiency of the EGFP site-specific knock-in could reach up to 29.6% at the pRosa26 locus in PFFs. Next, we used the EGFP reporter PFFs to address two key conditions in the process of achieving transgenic pigs, the limiting dilution method and the strategy to evaluate the safety and feasibility of the knock-in locus. This study demonstrates that we establish an efficient procedures for the exogenous gene knock-in technique and creates a platform to efficiently generate promoter-less and selectable marker-free transgenic PFFs through the CRISPR/Cas9 system. This study should contribute to the generation of promoter-less and selectable marker-free transgenic pigs and it may provide insights into sophisticated site-specific genome engineering techniques for additional species.

Homologous Recombination-Independent Large Gene Cassette Knock-in in CHO Cells Using TALEN and MMEJ-Directed Donor Plasmids

PubMed Central

Sakuma, Tetsushi; Takenaga, Mitsumasa; Kawabe, Yoshinori; Nakamura, Takahiro; Kamihira, Masamichi; Yamamoto, Takashi

2015-01-01

Gene knock-in techniques have rapidly evolved in recent years, along with the development and maturation of genome editing technology using programmable nucleases. We recently reported a novel strategy for microhomology-mediated end-joining-dependent integration of donor DNA by using TALEN or CRISPR/Cas9 and optimized targeting vectors, named PITCh (Precise Integration into Target Chromosome) vectors. Here we describe TALEN and PITCh vector-mediated integration of long gene cassettes, including a single-chain Fv-Fc (scFv-Fc) gene, in Chinese hamster ovary (CHO) cells, with comparison of targeting and cloning efficiency among several donor design and culture conditions. We achieved 9.6-kb whole plasmid integration and 7.6-kb backbone-free integration into a defined genomic locus in CHO cells. Furthermore, we confirmed the reasonable productivity of recombinant scFv-Fc protein of the knock-in cells. Using our protocol, the knock-in cell clones could be obtained by a single transfection and a single limiting dilution using a 96-well plate, without constructing targeting vectors containing long homology arms. Thus, the study described herein provides a highly practical strategy for gene knock-in of large DNA in CHO cells, which accelerates high-throughput generation of cell lines stably producing any desired biopharmaceuticals, including huge antibody proteins. PMID:26473830

Homologous Recombination-Independent Large Gene Cassette Knock-in in CHO Cells Using TALEN and MMEJ-Directed Donor Plasmids.

PubMed

Sakuma, Tetsushi; Takenaga, Mitsumasa; Kawabe, Yoshinori; Nakamura, Takahiro; Kamihira, Masamichi; Yamamoto, Takashi

2015-10-09

Gene knock-in techniques have rapidly evolved in recent years, along with the development and maturation of genome editing technology using programmable nucleases. We recently reported a novel strategy for microhomology-mediated end-joining-dependent integration of donor DNA by using TALEN or CRISPR/Cas9 and optimized targeting vectors, named PITCh (Precise Integration into Target Chromosome) vectors. Here we describe TALEN and PITCh vector-mediated integration of long gene cassettes, including a single-chain Fv-Fc (scFv-Fc) gene, in Chinese hamster ovary (CHO) cells, with comparison of targeting and cloning efficiency among several donor design and culture conditions. We achieved 9.6-kb whole plasmid integration and 7.6-kb backbone-free integration into a defined genomic locus in CHO cells. Furthermore, we confirmed the reasonable productivity of recombinant scFv-Fc protein of the knock-in cells. Using our protocol, the knock-in cell clones could be obtained by a single transfection and a single limiting dilution using a 96-well plate, without constructing targeting vectors containing long homology arms. Thus, the study described herein provides a highly practical strategy for gene knock-in of large DNA in CHO cells, which accelerates high-throughput generation of cell lines stably producing any desired biopharmaceuticals, including huge antibody proteins.

20 CFR 664.830 - How are performance measures for Youth Opportunity Grants determined?

Code of Federal Regulations, 2010 CFR

2010-04-01

... 20 Employees' Benefits 3 2010-04-01 2010-04-01 false How are performance measures for Youth Opportunity Grants determined? 664.830 Section 664.830 Employees' Benefits EMPLOYMENT AND TRAINING ADMINISTRATION, DEPARTMENT OF LABOR YOUTH ACTIVITIES UNDER TITLE I OF THE WORKFORCE INVESTMENT ACT Youth Opportunity Grants § 664.830 How are performance...

Effective Recreation Opportunity Set (EROS) Index: A Computable Measure of Recreation Supply

Treesearch

Donald B.K. English; H. Ken Cordell

1993-01-01

The EROS Set index is an economic measure of the abiliability of recreational opportunities to households and was developed for use in the 1989 Renewable Resources Planning Act Assessment. EROS indicies are loacation specific, and can be calcualted for any desired recreational enviorment or opportunity set using existing resource, population, and recreational trabel...

76 FR 76475 - Tregaron Opportunity Fund I, L.P.; Notice Seeking Exemption Under Section 312 of the Small...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-12-07

... SMALL BUSINESS ADMINISTRATION [License No. 09/09-0463] Tregaron Opportunity Fund I, L.P.; Notice Seeking Exemption Under Section 312 of the Small Business Investment Act, Conflicts of Interest Notice is hereby given that Tregaron Opportunity Fund I, L.P., 540 University Avenue, Suite 250, Palo Alto, CA...

75 FR 22436 - Notice With Respect to List of Countries Denying Fair Market Opportunities for Government-Funded...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-04-28

... market opportunities for products, suppliers or bidders of the United States in airport construction.... SUMMARY: Pursuant to section 533 of the Airport and Airway Improvement Act of 1982, as amended (49 U.S.C... denying fair market opportunities for U.S. products, suppliers, or bidders in foreign government-funded...

THE OPPORTUNITY TO LIVE IN DECENCY AND DIGNITY. ANNUAL REPORT, 1965.

ERIC Educational Resources Information Center

New Jersey Office of Economic Opportunity, Trenton.

THIS DOCUMENT IS THE REPORT OF THE FIRST YEAR OF NEW JERSEY'S ANTIPOVERTY EFFORTS FOLLOWING THE PASSAGE OF THE ECONOMIC OPPORTUNITY ACT OF 1964. DISCUSSED ARE COMMUNITY ACTION IN URBAN AND RURAL AREAS AND SUCH STATE PROGRAMS AS RURAL YOUTH DEVELOPMENT, MIGRANT OPPORTUNITIES, HEALTH SERVICES FOR POOR YOUTH, WORK EXPERIENCE FOR WELFARE RECIPIENTS,…

20 CFR 670.995 - What DOL equal opportunity and nondiscrimination regulations apply to Job Corps?

Code of Federal Regulations, 2010 CFR

2010-04-01

... TRAINING ADMINISTRATION, DEPARTMENT OF LABOR THE JOB CORPS UNDER TITLE I OF THE WORKFORCE INVESTMENT ACT Administrative and Management Provisions § 670.995 What DOL equal opportunity and nondiscrimination regulations...

Public Notice 2017-18: Seagis Property Group

EPA Pesticide Factsheets

EPA is providing notice of a proposed administrative penalty assessment for alleged violations of the Clean Water Act (Act). EPA is also providing notice of opportunity to comment on the proposed penalty assessment.

Public Notice 2017-21

EPA Pesticide Factsheets

The EPA is providing notice of a proposed administrative penalty assessment for alleged violations of the Clean Water Act (Act). The EPA is also providing notice of the opportunity to comment on the proposed penalty assessment.

Public Notice 2017-22

EPA Pesticide Factsheets

The EPA is providing notice of a proposed administrative penalty assessment for alleged violations of the Clean Water Act (Act). The EPA is also providing notice of the opportunity to comment on the proposed penalty assessment.

Public Notice 2017-25: Accurate Builders & Developers

EPA Pesticide Factsheets

EPA is providing notice of a proposed administrative penalty assessment for alleged violations of the Clean Water Act (Act). EPA is also providing notice of opportunity to comment on the proposed penalty assessment.

2018-09 - Kaplan Companies

EPA Pesticide Factsheets

EPA is providing notice of a proceeding to assess a Class II civil penalty for alleged violations of the Clean Water Act (Act). EPA is also providing notice of opportunity to comment on the proposed penalty assessment.

Learn about the EPA's Federal Technology Transfer Act Program

EPA Pesticide Factsheets

This act allows sharing Agency knowledge and expertise with outside partners through collaborative agreements and licensing. Potential partners can take advantage of opportunities to create or further develop solutions to environmental problems.

The new psychology of strategic leadership.

PubMed

Gavetti, Giovanni

2011-01-01

Firms in an industry typically cluster around a few strategic positions, and the intense competition on those occupied "mountaintops" makes it hard for firms to gain attractive returns. Superior opportunities lie on unoccupied mountaintops. Yet because those opportunities are "cognitively distant"--far from the status quo--strategists have trouble recognizing and acting on them. Competition, therefore, is weak. Most managers are trained to analyze economic forces when they want to identify new opportunities. But that approach usually won't uncover the kinds of ideas that overturn the status quo. Recent research on human cognition suggests that leaders would do better to use associative thinking to spot, act on, and legitimize distant opportunities. They should learn to make analogies with businesses in other industries, for example. For example, Charles Merrill launched an extraordinarily successful business when he reimagined banking as a "financial supermarket." This article explores ways to jump-start associational thinking--and to bring stakeholders along on the journey.

Cancer-associated isocitrate dehydrogenase 1 (IDH1) R132H mutation and d-2-hydroxyglutarate stimulate glutamine metabolism under hypoxia.

PubMed

Reitman, Zachary J; Duncan, Christopher G; Poteet, Ethan; Winters, Ali; Yan, Liang-Jun; Gooden, David M; Spasojevic, Ivan; Boros, Laszlo G; Yang, Shao-Hua; Yan, Hai

2014-08-22

Mutations in the cytosolic NADP(+)-dependent isocitrate dehydrogenase (IDH1) occur in several types of cancer, and altered cellular metabolism associated with IDH1 mutations presents unique therapeutic opportunities. By altering IDH1, these mutations target a critical step in reductive glutamine metabolism, the metabolic pathway that converts glutamine ultimately to acetyl-CoA for biosynthetic processes. While IDH1-mutated cells are sensitive to therapies that target glutamine metabolism, the effect of IDH1 mutations on reductive glutamine metabolism remains poorly understood. To explore this issue, we investigated the effect of a knock-in, single-codon IDH1-R132H mutation on the metabolism of the HCT116 colorectal adenocarcinoma cell line. Here we report the R132H-isobolome by using targeted (13)C isotopomer tracer fate analysis to trace the metabolic fate of glucose and glutamine in this system. We show that introduction of the R132H mutation into IDH1 up-regulates the contribution of glutamine to lipogenesis in hypoxia, but not in normoxia. Treatment of cells with a d-2-hydroxyglutarate (d-2HG) ester recapitulated these changes, indicating that the alterations observed in the knocked-in cells were mediated by d-2HG produced by the IDH1 mutant. These studies provide a dynamic mechanistic basis for metabolic alterations observed in IDH1-mutated tumors and uncover potential therapeutic targets in IDH1-mutated cancers. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

Conditional Sox9 ablation improves locomotor recovery after spinal cord injury by increasing reactive sprouting.

PubMed

McKillop, William M; York, Elisa M; Rubinger, Luc; Liu, Tony; Ossowski, Natalie M; Xu, Kathy; Hryciw, Todd; Brown, Arthur

2016-09-01

The absence of axonal regeneration after spinal cord injury (SCI) has been attributed to the up-regulation of axon-repelling molecules, such as chondroitin sulfate proteoglycans (CSPGs) present in the glial scar that forms post-SCI. We previously identified the transcription factor SOX9 as a key up-regulator of CSPG production and also demonstrated that conditional Sox9 ablation leads to decreased CSPG levels and improved recovery of hind limb function after SCI. We herein demonstrate increased neural input onto spinal neurons caudal to the lesion in spinal cord injured Sox9 conditional knock out mice as indicated by increased levels of the presynaptic markers synaptophysin and vesicular glutamate transporter 1 (VGLUT1) compared to controls. Axonal sparing, long-range axonal regeneration and reactive sprouting were investigated as possible explanations for the increase in neural inputs caudal to the lesion and for the improved locomotor outcomes in spinal cord-injured Sox9 conditional knock out mice. Whereas retrograde tract-tracing studies failed to reveal any evidence for increased axonal sparing or for long-range regeneration in the Sox9 conditional knock out mice, anterograde tract-tracing experiments demonstrated increased reactive sprouting caudal to the lesion after SCI. Finally we demonstrate that application of a broad spectrum MMP inhibitor to reduce CSPG degradation in Sox9 conditional knock out mice prevents the improvements in locomotor recovery observed in untreated Sox9 conditional knock out mice. These results suggest that improved recovery of locomotor function in Sox9 conditional knock out mice after SCI is due to increased reactive sprouting secondary to reduced CSPG levels distal to the lesion. Copyright © 2016 Elsevier Inc. All rights reserved.

Protein arginine methyltransferase 1 modulates innate immune responses through regulation of peroxisome proliferator-activated receptor γ-dependent macrophage differentiation.

PubMed

Tikhanovich, Irina; Zhao, Jie; Olson, Jody; Adams, Abby; Taylor, Ryan; Bridges, Brian; Marshall, Laurie; Roberts, Benjamin; Weinman, Steven A

2017-04-28

Arginine methylation is a common posttranslational modification that has been shown to regulate both gene expression and extranuclear signaling events. We recently reported defects in protein arginine methyltransferase 1 (PRMT1) activity and arginine methylation in the livers of cirrhosis patients with a history of recurrent infections. To examine the role of PRMT1 in innate immune responses in vivo , we created a cell type-specific knock-out mouse model. We showed that myeloid-specific PRMT1 knock-out mice demonstrate higher proinflammatory cytokine production and a lower survival rate after cecal ligation and puncture. We found that this defect is because of defective peroxisome proliferator-activated receptor γ (PPARγ)-dependent M2 macrophage differentiation. PPARγ is one of the key transcription factors regulating macrophage polarization toward a more anti-inflammatory and pro-resolving phenotype. We found that PRMT1 knock-out macrophages failed to up-regulate PPARγ expression in response to IL4 treatment resulting in 4-fold lower PPARγ expression in knock-out cells than in wild-type cells. Detailed study of the mechanism revealed that PRMT1 regulates PPARγ gene expression through histone H4R3me2a methylation at the PPARγ promoter. Supplementing with PPARγ agonists rosiglitazone and GW1929 was sufficient to restore M2 differentiation in vivo and in vitro and abrogated the difference in survival between wild-type and PRMT1 knock-out mice. Taken together these data suggest that PRMT1-dependent regulation of macrophage PPARγ expression contributes to the infection susceptibility in PRMT1 knock-out mice. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Starch Binding Domain-containing Protein 1 Plays a Dominant Role in Glycogen Transport to Lysosomes in Liver.

PubMed

Sun, Tao; Yi, Haiqing; Yang, Chunyu; Kishnani, Priya S; Sun, Baodong

2016-08-05

A small portion of cellular glycogen is transported to and degraded in lysosomes by acid α-glucosidase (GAA) in mammals, but it is unclear why and how glycogen is transported to the lysosomes. Stbd1 has recently been proposed to participate in glycogen trafficking to lysosomes. However, our previous study demonstrated that knockdown of Stbd1 in GAA knock-out mice did not alter lysosomal glycogen storage in skeletal muscles. To further determine whether Stbd1 participates in glycogen transport to lysosomes, we generated GAA/Stbd1 double knock-out mice. In fasted double knock-out mice, glycogen accumulation in skeletal and cardiac muscles was not affected, but glycogen content in liver was reduced by nearly 73% at 3 months of age and by 60% at 13 months as compared with GAA knock-out mice, indicating that the transport of glycogen to lysosomes was suppressed in liver by the loss of Stbd1. Exogenous expression of human Stbd1 in double knock-out mice restored the liver lysosomal glycogen content to the level of GAA knock-out mice, as did a mutant lacking the Atg8 family interacting motif (AIM) and another mutant that contains only the N-terminal 24 hydrophobic segment and the C-terminal starch binding domain (CBM20) interlinked by an HA tag. Our results demonstrate that Stbd1 plays a dominant role in glycogen transport to lysosomes in liver and that the N-terminal transmembrane region and the C-terminal CBM20 domain are critical for this function. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Protein phosphatase 2ACα gene knock-out results in cortical atrophy through activating hippo cascade in neuronal progenitor cells.

PubMed

Liu, Bo; Sun, Li-Hua; Huang, Yan-Fei; Guo, Li-Jun; Luo, Li-Shu

2018-02-01

Protein phosphatase 2ACα (PP2ACα), a vital member of the protein phosphatase family, has been studied primarily as a regulator for the development, growth and protein synthesis of a lot of cell types. Dysfunction of PP2ACα protein results in neurodegenerative disease; however, this finding has not been directly confirmed in the mouse model with PP2ACα gene knock-out. Therefore, in this study presented here, we generated the PP2ACα gene knock-out mouse model by the Cre-loxP targeting gene system, with the purpose to directly observe the regulatory role of PP2ACα gene in the development of mouse's cerebral cortex. We observe that knocking-out PP2ACα gene in the central nervous system (CNS) results in cortical neuronal shrinkage, synaptic plasticity impairments, and learning/memory deficits. Further study reveals that PP2ACα gene knock-out initiates Hippo cascade in cortical neuroprogenitor cells (NPCs), which blocks YAP translocation into the nuclei of NPCs. Notably, p73, directly targeted by Hippo cascade, can bind to the promoter of glutaminase2 (GLS2) that plays a dominant role in the enzymatic regulation of glutamate/glutamine cycle. Finally, we find that PP2ACα gene knock-out inhibits the glutamine synthesis through up-regulating the activity of phosphorylated-p73 in cortical NPCs. Taken together, it concludes that PP2ACα critically supports cortical neuronal growth and cognitive function via regulating the signaling transduction of Hippo-p73 cascade. And PP2ACα indirectly modulates the glutamine synthesis of cortical NPCs through targeting p73 that plays a direct transcriptional regulatory role in the gene expression of GLS2. Copyright © 2017 Elsevier Ltd. All rights reserved.

A Novel Mgp-Cre Knock-In Mouse Reveals an Anticalcification/Antistiffness Candidate Gene in the Trabecular Meshwork and Peripapillary Scleral Region.

PubMed

Borrás, Teresa; Smith, Matthew H; Buie, LaKisha K

2015-04-01

Soft tissue calcification is a pathological condition. Matrix Gla (MGP) is a potent mineralization inhibitor secreted by cartilage chondrocytes and arteries' vascular smooth muscle cells. Mgp knock-out mice die at 6 weeks due to massive arterial calcification. Arterial calcification results in arterial stiffness and higher systolic blood pressure. Intriguingly, MGP was highly abundant in trabecular meshwork (TM). Because tissue stiffness is relevant to glaucoma, we investigated which additional eye tissues use Mgp's function using knock-in mice. An Mgp-Cre-recombinase coding sequence (Cre) knock-in mouse, containing Mgp DNA plus an internal ribosomal entry site (IRES)-Cre-cassette was generated by homologous recombination. Founders were crossed with Cre-mediated reporter mouse R26R-lacZ. Their offspring expresses lacZ where Mgp is transcribed. Eyes from MgpCre/+;R26RlacZ/+ (Mgp-lacZ knock-in) and controls, 1 to 8 months were assayed for β-gal enzyme histochemistry. As expected, Mgp-lacZ knock-in's TM was intensely blue. In addition, this mouse revealed high specific expression in the sclera, particularly in the peripapillary scleral region (ppSC). Ciliary muscle and sclera above the TM were also positive. Scleral staining was located immediately underneath the choroid (chondrocyte layer), began midsclera and was remarkably high in the ppSC. Cornea, iris, lens, ciliary body, and retina were negative. All mice exhibited similar staining patterns. All controls were negative. Matrix Gla's restricted expression to glaucoma-associated tissues from anterior and posterior segments suggests its involvement in the development of the disease. Matrix Gla's anticalcification/antistiffness properties in the vascular tissue, together with its high TM and ppCS expression, place this gene as a strong candidate for TM's softness and sclera's stiffness regulation in glaucoma.

The kick-in system: a novel rapid knock-in strategy.

PubMed

Tomonoh, Yuko; Deshimaru, Masanobu; Araki, Kimi; Miyazaki, Yasuhiro; Arasaki, Tomoko; Tanaka, Yasuyoshi; Kitamura, Haruna; Mori, Fumiaki; Wakabayashi, Koichi; Yamashita, Sayaka; Saito, Ryo; Itoh, Masayuki; Uchida, Taku; Yamada, Junko; Migita, Keisuke; Ueno, Shinya; Kitaura, Hiroki; Kakita, Akiyoshi; Lossin, Christoph; Takano, Yukio; Hirose, Shinichi

2014-01-01

Knock-in mouse models have contributed tremendously to our understanding of human disorders. However, generation of knock-in animals requires a significant investment of time and effort. We addressed this problem by developing a novel knock-in system that circumvents several traditional challenges by establishing stem cells with acceptor elements enveloping a particular genomic target. Once established, these acceptor embryonic stem (ES) cells are efficient at directionally incorporating mutated target DNA using modified Cre/lox technology. This is advantageous, because knock-ins are not restricted to one a priori selected variation. Rather, it is possible to generate several mutant animal lines harboring desired alterations in the targeted area. Acceptor ES cell generation is the rate-limiting step, lasting approximately 2 months. Subsequent manipulations toward animal production require an additional 8 weeks, but this delimits the full period from conception of the genetic alteration to its animal incorporation. We call this system a "kick-in" to emphasize its unique characteristics of speed and convenience. To demonstrate the functionality of the kick-in methodology, we generated two mouse lines with separate mutant versions of the voltage-dependent potassium channel Kv7.2 (Kcnq2): p.Tyr284Cys (Y284C) and p.Ala306Thr (A306T); both variations have been associated with benign familial neonatal epilepsy. Adult mice homozygous for Y284C, heretofore unexamined in animals, presented with spontaneous seizures, whereas A306T homozygotes died early. Heterozygous mice of both lines showed increased sensitivity to pentylenetetrazole, possibly due to a reduction in M-current in CA1 hippocampal pyramidal neurons. Our observations for the A306T animals match those obtained with traditional knock-in technology, demonstrating that the kick-in system can readily generate mice bearing various mutations, making it a suitable feeder technology toward streamlined phenotyping.

Vapb/Amyotrophic lateral sclerosis 8 knock-in mice display slowly progressive motor behavior defects accompanying ER stress and autophagic response.

PubMed

Larroquette, Frédérique; Seto, Lesley; Gaub, Perrine L; Kamal, Brishna; Wallis, Deeann; Larivière, Roxanne; Vallée, Joanne; Robitaille, Richard; Tsuda, Hiroshi

2015-11-15

Missense mutations (P56S) in Vapb are associated with autosomal dominant motor neuron diseases: amyotrophic lateral sclerosis and lower motor neuron disease. Although transgenic mice overexpressing the mutant vesicle-associated membrane protein-associated protein B (VAPB) protein with neuron-specific promoters have provided some insight into the toxic properties of the mutant proteins, their role in pathogenesis remains unclear. To identify pathological defects in animals expressing the P56S mutant VAPB protein at physiological levels in the appropriate tissues, we have generated Vapb knock-in mice replacing wild-type Vapb gene with P56S mutant Vapb gene and analyzed the resulting pathological phenotypes. Heterozygous P56S Vapb knock-in mice show mild age-dependent defects in motor behaviors as characteristic features of the disease. The homozygous P56S Vapb knock-in mice show more severe defects compared with heterozygous mice reflecting the dominant and dose-dependent effects of P56S mutation. Significantly, the knock-in mice demonstrate accumulation of P56S VAPB protein and ubiquitinated proteins in cytoplasmic inclusions, selectively in motor neurons. The mutant mice demonstrate induction of ER stress and autophagic response in motor neurons before obvious onset of behavioral defects, suggesting that these cellular biological defects might contribute to the initiation of the disease. The P56S Vapb knock-in mice could be a valuable tool to gain a better understanding of the mechanisms by which the disease arises. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Generation of α-1,3-galactosyltransferase knocked-out transgenic cloned pigs with knocked-in five human genes.

PubMed

Kwon, Dae-Jin; Kim, Dong-Hwan; Hwang, In-Sul; Kim, Dong-Ern; Kim, Hyung-Joo; Kim, Jang-Seong; Lee, Kichoon; Im, Gi-Sun; Lee, Jeong-Woong; Hwang, Seongsoo

2017-02-01

Recent progress in genetic manipulation of pigs designated for xenotransplantation ha6s shown considerable promise on xenograft survival in primates. However, genetic modification of multiple genes in donor pigs by knock-out and knock-in technologies, aiming to enhance immunological tolerance against transplanted organs in the recipients, has not been evaluated for health issues of donor pigs. We produced transgenic Massachusetts General Hospital piglets by knocking-out the α-1,3-galactosyltransferase (GT) gene and by simultaneously knocking-in an expression cassette containing five different human genes including, DAF, CD39, TFPI, C1 inhibitor (C1-INH), and TNFAIP3 (A20) [GT -(DAF/CD39/TFPI/C1-INH/TNFAIP3)/+ ] that are connected by 2A peptide cleavage sequences to release individual proteins from a single translational product. All five individual protein products were successfully produced as determined by western blotting of umbilical cords from the newborn transgenic pigs. Although gross observation and histological examination revealed no significant pathological abnormality in transgenic piglets, hematological examination found that the transgenic piglets had abnormally low numbers of platelets and WBCs, including neutrophils, eosinophils, basophils, and lymphocytes. However, transgenic piglets had similar numbers of RBC and values of parameters related to RBC compared to the control littermate piglets. These data suggest that transgenic expression of those human genes in pigs impaired hematopoiesis except for erythropoiesis. In conclusion, our data suggest that transgenic expression of up to five different genes can be efficiently achieved and provide the basis for determining optimal dosages of transgene expression and combinations of the transgenes to warrant production of transgenic donor pigs without health issues.

Recovery Act. Tapoco project. Cheoah upgrade

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tran, Paul

2013-10-02

Under Funding Opportunity Announcement Number: DE-FOA-0000120, Recovery Act: Hydroelectric Facility Modernization, Alcoa Power Generating Inc. (APGI), a fully owned subsidiary of Alcoa Inc., implemented major upgrades at its Cheoah hydroelectric facility near Robbinsville, North Carolina.

Public Notice: 2018-09 Kaplan Companies

EPA Pesticide Factsheets

EPA is providing notice of a proceeding to assess a Class II civil penalty for alleged violations of the Clean Water Act (Act). EPA is also providing notice of opportunity to comment on the proposed penalty assessment.

Public Notice: 2018-10 Sun RB Development Company, LLC

EPA Pesticide Factsheets

EPA is providing notice of a proposed administrative penalty assessment for alleged violations of the Clean Water Act (Act). EPA is also providing notice of opportunity to comment on the proposed penalty assessment.

2017-21 Miller's Son Shine Acres, Inc.

EPA Pesticide Factsheets

The EPA is providing notice of a proposed administrative penalty assessment for alleged violations of the Clean Water Act (Act). The EPA is also providing notice of the opportunity to comment on the proposed penalty assessment.

Characterisation of a C1qtnf5 Ser163Arg Knock-In Mouse Model of Late-Onset Retinal Macular Degeneration

PubMed Central

Shu, Xinhua; Luhmann, Ulrich F. O.; Aleman, Tomas S.; Barker, Susan E.; Lennon, Alan; Tulloch, Brian; Chen, Mei; Xu, Heping; Jacobson, Samuel G.; Ali, Robin; Wright, Alan F.

2011-01-01

A single founder mutation resulting in a Ser163Arg substitution in the C1QTNF5 gene product causes autosomal dominant late-onset retinal macular degeneration (L-ORMD) in humans, which has clinical and pathological features resembling age-related macular degeneration. We generated and characterised a mouse “knock-in” model carrying the Ser163Arg mutation in the orthologous murine C1qtnf5 gene by site-directed mutagenesis and homologous recombination into mouse embryonic stem cells. Biochemical, immunological, electron microscopic, fundus autofluorescence, electroretinography and laser photocoagulation analyses were used to characterise the mouse model. Heterozygous and homozygous knock-in mice showed no significant abnormality in any of the above measures at time points up to 2 years. This result contrasts with another C1qtnf5 Ser163Arg knock-in mouse which showed most of the features of L-ORMD but differed in genetic background and targeting construct. PMID:22110650

A Study by High-Speed Photography of Combustion and Knock in a Spark-Ignition Engine

NASA Technical Reports Server (NTRS)

Miller, Cearcy D

1942-01-01

The study of combustion in a spark-ignition engine given in Technical Report no. 704 has been continued. The investigation was made with the NACA high-speed motion-picture camera and the NACA optical engine indicator. The camera operates at the rate of 40,000 photographs a second and makes possible the study of phenomena occurring in time intervals as short as 0.000025 second. Photographs are presented of combustion without knock and with both light and heavy knocks, the end zone of combustion being within the field of view. Time-pressure records covering the same conditions as the photographs are presented and their relations to the photographs are studied. Photographs with ignition at various advance angles are compared with a view to observing any possible relationship between pressure and flame depth. A tentative explanation of knock is suggested, which is designed to agree with the indications of the high-speed photographs and the time-pressure records.

Impaired fear extinction learning in adult heterozygous BDNF knock-out mice.

PubMed

Psotta, Laura; Lessmann, Volkmar; Endres, Thomas

2013-07-01

Brain-derived neurotrophic factor (BDNF) is a crucial regulator of neuroplasticity, which underlies learning and memory processes in different brain areas. To investigate the role of BDNF in the extinction of amygdala-dependent cued fear memories, we analyzed fear extinction learning in heterozygous BDNF knock-out mice, which possess a reduction of endogenous BDNF protein levels to ~50% of wild-type animals. Since BDNF expression has been shown to decline with aging of animals, we tested the performance in extinction learning of these mice at 2 months (young adults) and 7 months (older adults) of age. The present study shows that older adult heterozygous BDNF knock-out mice, which have a chronic 50% lack of BDNF, also possess a deficit in the acquisition of extinction memory, while extinction learning remains unaffected in young adult heterozygous BDNF knock-out mice. This deficit in extinction learning is accompanied by a reduction of BDNF protein in the hippocampus, amygdala and the prefrontal cortex. Copyright © 2013 Elsevier Inc. All rights reserved.

29 CFR 1626.12 - Conciliation efforts pursuant to section 7(d) of the Act.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 29 Labor 4 2012-07-01 2012-07-01 false Conciliation efforts pursuant to section 7(d) of the Act. 1626.12 Section 1626.12 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN EMPLOYMENT ACT § 1626.12 Conciliation efforts pursuant to section...

29 CFR 1626.21 - Effect of opinions and interpretations of the Commission.

Code of Federal Regulations, 2010 CFR

2010-07-01

.... 1626.21 Section 1626.21 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN EMPLOYMENT ACT § 1626.21 Effect of opinions and interpretations... Discrimination in Employment Act of 1967 through section 7(e)(1) of the Act, provides that: In any action or...

29 CFR 1626.12 - Conciliation efforts pursuant to section 7(d) of the Act.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 29 Labor 4 2014-07-01 2014-07-01 false Conciliation efforts pursuant to section 7(d) of the Act. 1626.12 Section 1626.12 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN EMPLOYMENT ACT § 1626.12 Conciliation efforts pursuant to section...

29 CFR 1626.21 - Effect of opinions and interpretations of the Commission.

Code of Federal Regulations, 2012 CFR

2012-07-01

.... 1626.21 Section 1626.21 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN EMPLOYMENT ACT § 1626.21 Effect of opinions and interpretations... Discrimination in Employment Act of 1967 through section 7(e)(1) of the Act, provides that: In any action or...

29 CFR 1626.12 - Conciliation efforts pursuant to section 7(d) of the Act.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 29 Labor 4 2010-07-01 2010-07-01 false Conciliation efforts pursuant to section 7(d) of the Act. 1626.12 Section 1626.12 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN EMPLOYMENT ACT § 1626.12 Conciliation efforts pursuant to section...

29 CFR 1626.12 - Conciliation efforts pursuant to section 7(d) of the Act.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 29 Labor 4 2013-07-01 2013-07-01 false Conciliation efforts pursuant to section 7(d) of the Act. 1626.12 Section 1626.12 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN EMPLOYMENT ACT § 1626.12 Conciliation efforts pursuant to section...

29 CFR 1626.21 - Effect of opinions and interpretations of the Commission.

Code of Federal Regulations, 2014 CFR

2014-07-01

.... 1626.21 Section 1626.21 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN EMPLOYMENT ACT § 1626.21 Effect of opinions and interpretations... Discrimination in Employment Act of 1967 through section 7(e)(1) of the Act, provides that: In any action or...

29 CFR 1626.12 - Conciliation efforts pursuant to section 7(d) of the Act.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 29 Labor 4 2011-07-01 2011-07-01 false Conciliation efforts pursuant to section 7(d) of the Act. 1626.12 Section 1626.12 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN EMPLOYMENT ACT § 1626.12 Conciliation efforts pursuant to section...

The Eilberg Act, New Seed Immigration, and Professional Labor Markets.

ERIC Educational Resources Information Center

Agarwal, Vinod B.; Yochum, Gilbert R.

1987-01-01

Foreign born professionals, especially those educated in the U.S., have become increasingly important to U.S. labor markets. This paper assesses the general effect of the Eilberg Act (1976) and subsequent U.S. Department of Labor implementations on new seed professionals' potential immigration patterns. The act could diminish opportunities for…

77 FR 58111 - Notice of Submission for OMB Review; Office of Postsecondary Education; Higher Education Act (HEA...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-09-19

...; Higher Education Act (HEA) Title II Report Cards on State Teacher Credentialing and Preparation SUMMARY: The Higher Education Opportunity Act of 2008 (HEOA) calls for annual reports from states and institutions of higher education (IHEs) on the quality of teacher preparation and state teacher certification...

76 FR 41263 - Notice of Intent To Award Affordable Care Act (ACA) Funding, EH10-1004

Federal Register 2010, 2011, 2012, 2013, 2014

2011-07-13

... Intent To Award Affordable Care Act (ACA) Funding, EH10-1004 Notice of Intent to award Affordable Care Act (ACA) funding to National Association for Public Health Statistics and Information Systems... under funding opportunity EH10-1004, ``National Environmental Public Health Tracking Program.'' AGENCY...

76 FR 59703 - Notice of Intent To Award Affordable Care Act (ACA) Funding, RFA-TP-08-001

Federal Register 2010, 2011, 2012, 2013, 2014

2011-09-27

... Intent To Award Affordable Care Act (ACA) Funding, RFA- TP-08-001 AGENCY: Centers for Disease Control and... of Intent to award Affordable Care Act (ACA) funding to Preparedness and Emergency Response Research... continuation application under Funding Opportunity Announcement RFA-TP- 08-001, ``Preparedness and Emergency...

77 FR 2265 - Request for Extension and Revision of a Currently Approved Information Collection

Federal Register 2010, 2011, 2012, 2013, 2014

2012-01-17

... States Grain Standards Act and the Agricultural Marketing Act of 1946. This voluntary survey gives... the Agricultural Marketing Act (AMA) (7 U.S.C. 1621-1627) to facilitate the marketing of grain... agricultural commodity markets an opportunity to provide feedback on the quality of services they receive and...

29 CFR 34.1 - Purpose; application.

Code of Federal Regulations, 2010 CFR

2010-07-01

... Assistance Act of 1974, as amended (38 U.S.C. 4212), the Equal Pay Act of 1963, as amended (29 U.S.C. 206d... Secretary of Labor IMPLEMENTATION OF THE NONDISCRIMINATION AND EQUAL OPPORTUNITY REQUIREMENTS OF THE JOB TRAINING PARTNERSHIP ACT OF 1982, AS AMENDED (JTPA) General Provisions § 34.1 Purpose; application. (a...

29 CFR 34.12 - Delegation and coordination.

Code of Federal Regulations, 2010 CFR

2010-07-01

... amended (38 U.S.C. 4212), the Equal Pay Act of 1963, as amended, title VII of the Civil Rights Act of 1964... Office of the Secretary of Labor IMPLEMENTATION OF THE NONDISCRIMINATION AND EQUAL OPPORTUNITY REQUIREMENTS OF THE JOB TRAINING PARTNERSHIP ACT OF 1982, AS AMENDED (JTPA) General Provisions § 34.12...

Bright New Era in Education.

ERIC Educational Resources Information Center

Department of Education, Washington, DC.

During 1993-94, the U.S. Congress passed several initiatives that offer great potential for facilitating progress in education. These include: (1) The Goals 2000: Educate America Act; (2) Individual Education Accounts; (3) the Family Involvement Initiative; (4) the Safe Schools Act (as part of Goals 2000); (5) the School-to-Work Opportunities Act;…

The Every Student Succeeds Act: Opportunities and Responsibilities. re:VISION

ERIC Educational Resources Information Center

Johnson, Mark

2016-01-01

The "Every Student Succeeds Act" (ESSA) was signed into law by President Barack Obama on December 10, 2015. ESSA reauthorizes the "Elementary and Secondary Education Act" (ESEA), which was first enacted as part of President Lyndon B. Johnson's "War on Poverty." Since 1965, ESEA has represented the federal government's…

Students with Disabilities at Degree-Granting Postsecondary Institutions. First Look. NCES 2011-018

ERIC Educational Resources Information Center

Raue, Kimberley; Lewis, Laurie

2011-01-01

Growing enrollments of students with disabilities in postsecondary education along with recent key legislation such as the Americans with Disabilities Act Amendments Act of 2008 and the 2008 Higher Education Opportunity Act, have generated considerable interest in research on accessibility of higher education for students with disabilities. This…

20 CFR 664.820 - Who is eligible to receive services under Youth Opportunity Grants?

Code of Federal Regulations, 2010 CFR

2010-04-01

... 20 Employees' Benefits 3 2010-04-01 2010-04-01 false Who is eligible to receive services under Youth Opportunity Grants? 664.820 Section 664.820 Employees' Benefits EMPLOYMENT AND TRAINING ADMINISTRATION, DEPARTMENT OF LABOR YOUTH ACTIVITIES UNDER TITLE I OF THE WORKFORCE INVESTMENT ACT Youth Opportunity Grants § 664.820 Who is eligible to...

20 CFR 664.600 - Are Local Boards required to offer summer employment opportunities in the local youth program?

Code of Federal Regulations, 2012 CFR

2012-04-01

... 20 Employees' Benefits 4 2012-04-01 2012-04-01 false Are Local Boards required to offer summer... WORKFORCE INVESTMENT ACT Summer Employment Opportunities § 664.600 Are Local Boards required to offer summer employment opportunities in the local youth program? (a) Yes, Local Boards are required to offer summer youth...

20 CFR 664.600 - Are Local Boards required to offer summer employment opportunities in the local youth program?

Code of Federal Regulations, 2011 CFR

2011-04-01

... 20 Employees' Benefits 3 2011-04-01 2011-04-01 false Are Local Boards required to offer summer... INVESTMENT ACT Summer Employment Opportunities § 664.600 Are Local Boards required to offer summer employment opportunities in the local youth program? (a) Yes, Local Boards are required to offer summer youth employment...

20 CFR 664.600 - Are Local Boards required to offer summer employment opportunities in the local youth program?

Code of Federal Regulations, 2014 CFR

2014-04-01

... 20 Employees' Benefits 4 2014-04-01 2014-04-01 false Are Local Boards required to offer summer... WORKFORCE INVESTMENT ACT Summer Employment Opportunities § 664.600 Are Local Boards required to offer summer employment opportunities in the local youth program? (a) Yes, Local Boards are required to offer summer youth...

20 CFR 664.600 - Are Local Boards required to offer summer employment opportunities in the local youth program?

Code of Federal Regulations, 2013 CFR

2013-04-01

... 20 Employees' Benefits 4 2013-04-01 2013-04-01 false Are Local Boards required to offer summer... WORKFORCE INVESTMENT ACT Summer Employment Opportunities § 664.600 Are Local Boards required to offer summer employment opportunities in the local youth program? (a) Yes, Local Boards are required to offer summer youth...

Evaluation of the DC Opportunity Scholarship Program: Impacts after One Year. NCEE 2017-4022

ERIC Educational Resources Information Center

Dynarski, Mark; Rui, Ning; Webber, Ann; Gutmann, Babette

2017-01-01

The District of Columbia Opportunity Scholarship Program (OSP) was created by Congress to provide tuition vouchers to low-income parents who want their child to attend a private school. The Scholarships for Opportunity and Results (SOAR) Act of 2011 also mandated an evaluation of the OSP program. This report examines impacts one year after…

78 FR 17705 - Privacy Act of 1974, as Amended; Notice of a New System of Records

Federal Register 2010, 2011, 2012, 2013, 2014

2013-03-22

..., awarding scholarships and identifying educational opportunities, promoting business opportunities... responsibilities of OIA, including promoting economic development initiatives and employment and educational... Form number and associated data, educational history, and professional licensing information. Many...

Species specific inhibition of viral replication using dicer substrate siRNAs (DsiRNAs) targeting the viral nucleoprotein of the fish pathogenic rhabdovirus viral hemorrhagic septicemia virus (VHSV).

PubMed

Bohle, Harry; Lorenzen, Niels; Schyth, Brian Dall

2011-06-01

Gene knock down by the use of small interfering RNAs (siRNAs) is widely used as a method for reducing the expression of specific genes in eukaryotic cells via the RNA interference pathway. But, the effectivity of siRNA induced gene knock down in cells from fish has in several studies been questioned and the specificity seems to be a general problem in cells originating from both lower and higher vertebrates. Here we show that we are able to reduce the level of viral gene expression and replication specifically in fish cells in vitro. We do so by using 27/25-mer DsiRNAs acting as substrates for dicer for the generation of siRNAs targeting the nucleoprotein N gene of viral hemorrhagic septicemia virus (VHSV). This rhabdovirus infects salmonid fish and is responsible for large yearly losses in aquaculture production. Specificity of the DsiRNA is assured in two ways: first, by using the conventional method of testing a control DsiRNA which should not target the gene of interest. Second, by assuring that replication of a heterologous virus of the same genus as the target virus was not inhibited by the DsiRNA. Target controls are, as we have previously highlighted, essential for verification of the specificity of siRNA-induced interference with virus multiplication, but they are still not in general use. Copyright © 2011 Elsevier B.V. All rights reserved.

Stable knock-down of the sphingosine 1-phosphate receptor S1P1 influences multiple functions of human endothelial cells.

PubMed

Krump-Konvalinkova, Vera; Yasuda, Satoshi; Rubic, Tina; Makarova, Natalia; Mages, Jörg; Erl, Wolfgang; Vosseler, Claudia; Kirkpatrick, C James; Tigyi, Gabor; Siess, Wolfgang

2005-03-01

Sphingosine 1-phosphate (S1P) is a bioactive phospholipid acting both as a ligand for the G protein-coupled receptors S1P1-5 and as a second messenger. Because S1P1 knockout is lethal in the transgenic mouse, an alternative approach to study the function of S1P1 in endothelial cells is needed. All human endothelial cells analyzed expressed abundant S1P1 transcripts. We permanently silenced (by RNA interference) the expression of S1P1 in the human endothelial cell lines AS-M.5 and ISO-HAS.1. The S1P1 knock-down cells manifested a distinct morphology and showed neither actin ruffles in response to S1P nor an angiogenic reaction. In addition, these cells were more sensitive to oxidant stress-mediated injury. New S1P1-dependent gene targets were identified in human endothelial cells. S1P1 silencing decreased the expression of platelet-endothelial cell adhesion molecule-1 and VE-cadherin and abolished the induction of E-selectin after cell stimulation with lipopolysaccharide or tumor necrosis factor-alpha. Microarray analysis revealed downregulation of further endothelial specific transcripts after S1P1 silencing. Long-term silencing of S1P1 enabled us for the first time to demonstrate the involvement of S1P1 in key functions of endothelial cells and to identify new S1P1-dependent gene targets.

Impact of Formaldehyde Addition on Auto-Ignition in Internal-Combustion Engines

NASA Astrophysics Data System (ADS)

Kuwahara, Kazunari; Ando, Hiromitsu; Furutani, Masahiro; Ohta, Yasuhiko

By employing a direct-injection diesel engine equipped with a common-rail type of injection system, by adding formaldehyde (CH2O) to the intake air, and by changing the fuel-injection timing, the compression ratio and the intake-air temperature, a mechanism for CH2O as a fuel additive to affect auto-ignition was discussed. Unlike an HCCI type of engine, the diesel engine can expose an air-fuel mixture only to a limited range of the in-cylinder temperature before the ignition, and can separate low- and high-temperature parts of the mechanism. When low-temperature oxidation starts at a temperature above 900K, there are cases that the CH2O advances the ignition timing. Below 900K, to the contrary, it always retards the timing. It is because, above 900K, a part of the CH2O changes into CO together with H2O2 as an ignition promoter. Below 900K, on the other hand, the CH2O itself acts as an OH radical scavenger against cool-flame reaction, from the beginning of low-temperature oxidation. Then, the engine was modified for its extraordinary function as a gasoline-knocking generator, in order that an effect of CH2O on knocking could be discussed. The CH2O retards the onset of auto-ignition of an end gas. Judging from a large degree of the retardation, the ignition is probably triggered below 900K.

A survey of transportation planning practices in state departments of transportation.

DOT National Transportation Integrated Search

1999-03-01

The Intermodal Surface Transportation Efficiency Act (ISTEA), and now its successor the Transportation Equity Act for the 21st Century (TEA-21), fundamentally altered the transportation planning process, providing new opportunities and new challenges...

Public Notice 2018-8 City of Peekskill

EPA Pesticide Factsheets

EPA is providing notice of a proceeding to assess a Class I civil penalty for alleged violations of the Clean Water Act (Act). EPA is also providing notice of opportunity to comment on the proposed penalty assessment.

PDE5 Inhibitors Enhance Celecoxib Killing in Multiple Tumor Types

PubMed Central

BOOTH, LAURENCE; ROBERTS, JANE L.; CRUICKSHANKS, NICHOLA; TAVALLAI, SEYEDMEHRAD; WEBB, TIMOTHY; SAMUEL, PETER; CONLEY, ADAM; BINION, BRITTANY; YOUNG, HAROLD F.; POKLEPOVIC, ANDREW; SPIEGEL, SARAH; DENT, PAUL

2015-01-01

The present studies determined whether clinically relevant phosphodiesterase 5 (PDE5) inhibitors interacted with a clinically relevant NSAID, celecoxib, to kill tumor cells. Celecoxib and PDE5 inhibitors interacted in a greater than additive fashion to kill multiple tumor cell types. Celecoxib and sildenafil killed ex vivo primary human glioma cells as well as their associated activated microglia. Knock down of PDE5 recapitulated the effects of PDE5 inhibitor treatment; the nitric oxide synthase inhibitor L-NAME suppressed drug combination toxicity. The effects of celecoxib were COX2 independent. Over-expression of c-FLIP-s or knock down of CD95/FADD significantly reduced killing by the drug combination. CD95 activation was dependent on nitric oxide and ceramide signaling. CD95 signaling activated the JNK pathway and inhibition of JNK suppressed cell killing. The drug combination inactivated mTOR and increased the levels of autophagy and knock down of Beclin1 or ATG5 strongly suppressed killing by the drug combination. The drug combination caused an ER stress response; knock down of IRE1α/XBP1 enhanced killing whereas knock down of eIF2α/ATF4/CHOP suppressed killing. Sildenafil and celecoxib treatment suppressed the growth of mammary tumors in vivo. Collectively our data demonstrate that clinically achievable concentrations of celecoxib and sildenafil have the potential to be a new therapeutic approach for cancer. PMID:25303541

The Brain Proteome of the Ubiquitin Ligase Peli1 Knock-Out Mouse during Experimental Autoimmune Encephalomyelitis.

PubMed

Lereim, Ragnhild Reehorst; Oveland, Eystein; Xiao, Yichuan; Torkildsen, Øivind; Wergeland, Stig; Myhr, Kjell-Morten; Sun, Shao-Cong; Berven, Frode S

2016-09-01

The ubiquitin ligase Peli1 has previously been suggested as a potential treatment target in multiple sclerosis. In the multiple sclerosis disease model, experimental autoimmune encephalomyelitis, Peli1 knock-out led to less activated microglia and less inflammation in the central nervous system. Despite being important in microglia, Peli1 expression has also been detected in glial and neuronal cells. In the present study the overall brain proteomes of Peli1 knock-out mice and wild-type mice were compared prior to experimental autoimmune encephalomyelitis induction, at onset of the disease and at disease peak. Brain samples from the frontal hemisphere, peripheral from the extensive inflammatory foci, were analyzed using TMT-labeling of sample pools, and the discovered proteins were verified in individual mice using label-free proteomics. The greatest proteomic differences between Peli1 knock-out and wild-type mice were observed at the disease peak. In Peli1 knock-out a higher degree of antigen presentation, increased activity of adaptive and innate immune cells and alterations to proteins involved in iron metabolism were observed during experimental autoimmune encephalomyelitis. These results unravel global effects to the brain proteome when abrogating Peli1 expression, underlining the importance of Peli1 as a regulator of the immune response also peripheral to inflammatory foci during experimental autoimmune encephalomyelitis. The proteomics data is available in PRIDE with accession PXD003710.

Genome editing using CRISPR/Cas9-based knock-in approaches in zebrafish.

PubMed

Albadri, Shahad; Del Bene, Filippo; Revenu, Céline

2017-05-15

With its variety of applications, the CRISPR/Cas9 genome editing technology has been rapidly evolving in the last few years. In the zebrafish community, knock-out reports are constantly increasing but insertion studies have been so far more challenging. With this review, we aim at giving an overview of the homologous directed repair (HDR)-based knock-in generation in zebrafish. We address the critical points and limitations of the procedure such as cutting efficiency of the chosen single guide RNA, use of cas9 mRNA or Cas9 protein, homology arm size etc. but also ways to circumvent encountered issues with HDR insertions by the development of non-homologous dependent strategies. While imprecise, these homology-independent mechanisms based on non-homologous-end-joining (NHEJ) repair have been employed in zebrafish to generate reporter lines or to accurately edit an open reading frame by the use of intron-targeting modifications. Therefore, with higher efficiency and insertion rate, NHEJ-based knock-in seems to be a promising approach to target endogenous loci and to circumvent the limitations of HDR whenever it is possible and appropriate. In this perspective, we propose new strategies to generate cDNA edited or tagged insertions, which once established will constitute a new and versatile toolbox for CRISPR/Cas9-based knock-ins in zebrafish. Copyright © 2017 Elsevier Inc. All rights reserved.

Alternative funding opportunities for National Park Service Transit

DOT National Transportation Integrated Search

2014-01-01

The Moving Ahead for Progress in the 21st Century Act and other anticipated financial changes are expected to reduce funding for National Park Service (NPS) transit systems. This paper discusses opportunities that may exist for NPS to work with the F...

28 CFR 42.301 - Purpose.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 28 Judicial Administration 1 2010-07-01 2010-07-01 false Purpose. 42.301 Section 42.301 Judicial Administration DEPARTMENT OF JUSTICE NONDISCRIMINATION; EQUAL EMPLOYMENT OPPORTUNITY; POLICIES AND PROCEDURES... opportunities in the criminal justice system is a necessary component to the Safe Streets Act's program to...

Savvy Money Management Can Soothe the Sting of the Arbitrage Rebate.

ERIC Educational Resources Information Center

Resnick, Michael A.

1988-01-01

For school systems, the Tax Reform Act of 1986 increased the relative cost of borrowing and put greater limits on investment opportunities. Offers tips to help school boards ask the right questions to take advantage of all possible opportunities. (MLF)

40 CFR 745.227 - Work practice standards for conducting lead-based paint activities: target housing and child...

Code of Federal Regulations, 2011 CFR

2011-07-01

... impact surface that is cause by impact from a related building component (such as a door knob that knocks into a wall or a door that knocks against its door frame; and (iv) If there is any other deteriorated...

40 CFR 745.227 - Work practice standards for conducting lead-based paint activities: target housing and child...

Code of Federal Regulations, 2010 CFR

2010-07-01

... impact surface that is cause by impact from a related building component (such as a door knob that knocks into a wall or a door that knocks against its door frame; and (iv) If there is any other deteriorated...

The Clean Air Act and the Economy

EPA Pesticide Factsheets

Since 1970, cleaner air and a growing economy have gone hand in hand. The Act has created market opportunities that have helped to inspire innovation in cleaner technologies for which the United States has become a global market leader.

School-to-Work Opportunities for All Youth: Intermediary Organizations and Expanding Options. School-to-Work Intermediary Project. Issue Brief.

ERIC Educational Resources Information Center

Spera, Vincent; Williams, Andra

The School to Work Opportunities Act of 1994 requires that all young people, including young people with disabilities and out-of-school youth, have equal opportunities to participate in the activities funded by it. Local intermediary organizations that provide a critical convening role and offer services to core education and business partners can…

Community health workers and the Patient Protection and Affordable Care Act: an opportunity for a research, advocacy, and policy agenda.

PubMed

Shah, Megha K; Heisler, Michele; Davis, Matthew M

2014-02-01

Community health workers (CHWs), who have been shown to be effective in multiple roles in the provision of culturally appropriate health care in a variety of settings, have the potential to be important members of an interdisciplinary health care team. Recent efforts have started to explore how best to integrate CHWs into the health system. However, to date, there has been limited policy guidance, support, or evidence on how best to achieve this on a larger scale. The Patient Protection and Affordable Care Act (ACA), through several provisions, provides a unique opportunity to create a unified framework for workforce integration and wider utilization of CHWs. This review identifies four major opportunities to further the research, advocacy, and policy agenda for CHWs.

29 CFR 1626.2 - Terms defined in the Age Discrimination in Employment Act of 1967, as amended.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 29 Labor 4 2010-07-01 2010-07-01 false Terms defined in the Age Discrimination in Employment Act of 1967, as amended. 1626.2 Section 1626.2 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN EMPLOYMENT ACT § 1626.2 Terms defined in...

29 CFR 1626.2 - Terms defined in the Age Discrimination in Employment Act of 1967, as amended.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 29 Labor 4 2012-07-01 2012-07-01 false Terms defined in the Age Discrimination in Employment Act of 1967, as amended. 1626.2 Section 1626.2 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN EMPLOYMENT ACT § 1626.2 Terms defined in...

29 CFR 1604.8 - Relationship of title VII to the Equal Pay Act.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 29 Labor 4 2010-07-01 2010-07-01 false Relationship of title VII to the Equal Pay Act. 1604.8 Section 1604.8 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION GUIDELINES ON DISCRIMINATION BECAUSE OF SEX § 1604.8 Relationship of title VII to the Equal Pay Act. (a) The...

29 CFR 1626.2 - Terms defined in the Age Discrimination in Employment Act of 1967, as amended.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 29 Labor 4 2013-07-01 2013-07-01 false Terms defined in the Age Discrimination in Employment Act of 1967, as amended. 1626.2 Section 1626.2 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN EMPLOYMENT ACT § 1626.2 Terms defined in...

29 CFR 1604.8 - Relationship of title VII to the Equal Pay Act.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 29 Labor 4 2012-07-01 2012-07-01 false Relationship of title VII to the Equal Pay Act. 1604.8 Section 1604.8 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION GUIDELINES ON DISCRIMINATION BECAUSE OF SEX § 1604.8 Relationship of title VII to the Equal Pay Act. (a) The...

29 CFR 1626.2 - Terms defined in the Age Discrimination in Employment Act of 1967, as amended.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 29 Labor 4 2011-07-01 2011-07-01 false Terms defined in the Age Discrimination in Employment Act of 1967, as amended. 1626.2 Section 1626.2 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN EMPLOYMENT ACT § 1626.2 Terms defined in...

29 CFR 1604.8 - Relationship of title VII to the Equal Pay Act.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 29 Labor 4 2013-07-01 2013-07-01 false Relationship of title VII to the Equal Pay Act. 1604.8 Section 1604.8 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION GUIDELINES ON DISCRIMINATION BECAUSE OF SEX § 1604.8 Relationship of title VII to the Equal Pay Act. (a) The...

29 CFR 1604.8 - Relationship of title VII to the Equal Pay Act.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 29 Labor 4 2011-07-01 2011-07-01 false Relationship of title VII to the Equal Pay Act. 1604.8 Section 1604.8 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION GUIDELINES ON DISCRIMINATION BECAUSE OF SEX § 1604.8 Relationship of title VII to the Equal Pay Act. (a) The...

29 CFR 1626.2 - Terms defined in the Age Discrimination in Employment Act of 1967, as amended.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 29 Labor 4 2014-07-01 2014-07-01 false Terms defined in the Age Discrimination in Employment Act of 1967, as amended. 1626.2 Section 1626.2 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN EMPLOYMENT ACT § 1626.2 Terms defined in...

The Adult Education Act 1964-1979: A Political History.

ERIC Educational Resources Information Center

DeSanctis, Vincent

The Adult Education Act, which provides federal funds for adult basic education (ABE) programs, has undergone many changes since enactment in 1964. Passage of ABE legislation as Title 11B under the Office of Economic Opportunity Act was a political solution for adult educators who had sought unsuccessfully for years for federal ABE legislation.…

12 CFR 268.503 - Enforcement of final EEOC decisions.

Code of Federal Regulations, 2010 CFR

2010-01-01

... the decision pursuant to title VII, the ADEA, the Equal Pay Act or the Rehabilitation Act and to seek... RESERVE SYSTEM RULES REGARDING EQUAL OPPORTUNITY Remedies and Enforcement § 268.503 Enforcement of final... Procedures Act, 5 U.S.C. 701 et seq., and the mandamus statute, 28 U.S.C. 1361, or to commence de novo...

Self-Delivering RNAi Targeting PD-1 Improves Tumor-Specific T Cell Functionality for Adoptive Cell Therapy of Malignant Melanoma.

PubMed

Ligtenberg, Maarten A; Pico de Coaña, Yago; Shmushkovich, Taisia; Yoshimoto, Yuya; Truxova, Iva; Yang, Yuan; Betancur-Boissel, Monica; Eliseev, Alexey V; Wolfson, Alexey D; Kiessling, Rolf

2018-06-06

Adoptive cell therapy (ACT) is becoming a prominent alternative therapeutic treatment for cancer patients relapsing on traditional therapies. In parallel, antibodies targeting immune checkpoint molecules, such as cytotoxic-T-lymphocyte-associated antigen 4 (CTLA-4) and cell death protein 1 pathway (PD-1), are rapidly being approved for multiple cancer types, including as first line therapy for PD-L1-expressing non-small-cell lung cancer. The combination of ACT and checkpoint blockade could substantially boost the efficacy of ACT. In this study, we generated a novel self-delivering small interfering RNA (siRNA) (sdRNA) that knocked down PD-1 expression on healthy donor T cells as well as patient-derived tumor-infiltrating lymphocytes (TIL). We have developed an alternative chemical modification of RNA backbone for improved stability and increased efficacy. Our results show that T cells treated with sdRNA specific for PD-1 had increased interferon γ (IFN-γ) secreting capacity and that this modality of gene expression interference could be utilized in our rapid expansion protocol for production of TIL for therapy. TIL expanded in the presence of PD-1-specific sdRNA performed with increased functionality against autologous tumor as compared to control TIL. This method of introducing RNAi into T cells to modify the expression of proteins could easily be adopted into any ACT protocol and will lead to the exploration of new combination therapies. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

29 CFR 1626.3 - Other definitions.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 29 Labor 4 2014-07-01 2014-07-01 false Other definitions. 1626.3 Section 1626.3 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN... Discrimination in Employment Act of 1967, as amended; the Commission shall mean the Equal Employment Opportunity...

29 CFR 1626.3 - Other definitions.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 29 Labor 4 2012-07-01 2012-07-01 false Other definitions. 1626.3 Section 1626.3 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN... Discrimination in Employment Act of 1967, as amended; the Commission shall mean the Equal Employment Opportunity...

29 CFR 1626.3 - Other definitions.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 29 Labor 4 2013-07-01 2013-07-01 false Other definitions. 1626.3 Section 1626.3 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN... Discrimination in Employment Act of 1967, as amended; the Commission shall mean the Equal Employment Opportunity...

29 CFR 1626.3 - Other definitions.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 29 Labor 4 2011-07-01 2011-07-01 false Other definitions. 1626.3 Section 1626.3 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN... Discrimination in Employment Act of 1967, as amended; the Commission shall mean the Equal Employment Opportunity...

29 CFR 1626.3 - Other definitions.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 29 Labor 4 2010-07-01 2010-07-01 false Other definitions. 1626.3 Section 1626.3 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN... Discrimination in Employment Act of 1967, as amended; the Commission shall mean the Equal Employment Opportunity...

A Lifetime of Learning

ERIC Educational Resources Information Center

Office of Education (DHEW), Washington, DC.

This booklet discusses three Federal programs for the continuing education of adults: Adult Basic Education; Community Service and Continuing Education; and Civil Defense Education. Initiated through the Economic Opportunity Act of 1964, Adult Basic Education began operation in 1965, financed by the Office of Economic Opportunity and administered…

Application of systems thinking in health: opportunities for translating theory into practice Comment on "Constraints to applying systems thinking concepts in health systems: a regional perspective from surveying stakeholders in Eastern Mediterranean countries".

PubMed

Malik, Asmat Ullah

2015-03-19

Systems thinking is not a new concept to health system strengthening; however, one question remains unanswered: How policy-makers, system designers and consultants with a system thinking philosophy should act (have acted) as potential change agents in actually gaining opportunities to introduce systems thinking? Development of Comprehensive Multi-Year Plans (cMYPs) for Immunization System is one such opportunity because almost all Low- and Middle-Income Countries (LMICs) develop and implement cMYPs every five years. Without building upon examples and showing practical application, the discussions and deliberations on systems thinking may fade away with passage of time. There are opportunities that exist around us in our existing health systems that we can benefit from starting with an incremental approach and generating evidence for longer lasting system-wide changes. © 2015 by Kerman University of Medical Sciences.

To amend the Interstate Land Sales Full Disclosure Act to clarify how the Act applies to condominiums.

THOMAS, 112th Congress

Rep. Maloney, Carolyn B. [D-NY-14

2012-08-02

House - 10/01/2012 Referred to the Subcommittee on Insurance, Housing and Community Opportunity. (All Actions) Tracker: This bill has the status IntroducedHere are the steps for Status of Legislation:

Pesky Personnel Paperwork: Federal Reporting Requirements.

ERIC Educational Resources Information Center

Guare, Timothy E.; Bush, Robert K.

1994-01-01

Identifies federal reporting obligations that govern school districts' human resources management in the following areas: (1) Internal Revenue Service filing requirements; (2) Fair Labor Standards Act recordkeeping; (3) Equal Employment Opportunity Commission reporting; and (4) Family and Medical Leave Act recordkeeping. (MLF)

76 FR 58045 - Notice of Determinations Regarding Eligibility To Apply for Worker Adjustment Assistance and...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-09-19

... the Andean Trade Preference Act, African Growth and Opportunity Act, or the Caribbean Basin Economic... more than one certification at a time. TA-W-80,293; Klaussner Furniture Industries, Milford, IA TA-W-80...

Is There a Mobile Social Presence?

ERIC Educational Resources Information Center

Tu, Chih-Hsiung; McIsaac, Marina; Sujo-Montes, Laura; Armfield, Shadow

2012-01-01

Mobile learning environments are human networks that afford the opportunity to participate in creative endeavors, social networking, organize/reorganize social contents, and manage social acts at anytime, anywhere through mobile technologies. Social acts that elicit identities, develop awareness, cement relationships, ensure connections, and…

Nontraditional Employment Policy Package.

ERIC Educational Resources Information Center

Wider Opportunities for Women, Inc., Washington, DC.

This document contains the text and summaries of provisions, along with position papers on four pieces of federal legislation affecting nontraditional job opportunities for women: (1) Nontraditional Employment of Women Act; (2) Women in Apprenticeship Occupations and Nontraditional Occupations Act; (3) Intermodal Surface Transportation Efficiency…

State Implementation of Reforms Promoted under the Recovery Act. A Report from Charting the Progress of Education Reform: An Evaluation of the Recovery Act's Role. NCEE 2014-4011

ERIC Educational Resources Information Center

Webber, Ann; Troppe, Patricia; Milanowski, Anthony; Gutmann, Babette; Reisner, Elizabeth; Goertz, Margaret

2014-01-01

The American Recovery and Reinvestment Act (ARRA or the Recovery Act) of 2009 provided an unprecedented level of funding for K-12 education. The program created a "historic opportunity to save hundreds of thousands of jobs, support states and school districts, and advance reforms and improvements that will create long-lasting results for our…

Better Evidence, Better Choices, Better Schools: State Supports for Evidence-Based School Improvement and the Every Student Succeeds Act. Series on Implementation of the Every Student Succeeds Act

ERIC Educational Resources Information Center

Fleischman, Steve; Scott, Caitlin; Sargrad, Scott

2016-01-01

Signed into law in December 2015, the Every Student Suceeds Act (ESSA) offers state education agencies significant opportunities to use evidence to support the improvement of schools and ensure better outcomes for all students. ESSA replaces the law, regulations, and guidance established through the 2001 No Child Left Behind Act (NCLB), but two…

50th Anniversary of the Civil Rights Act of 1964

NASA Image and Video Library

2014-06-23

From left: Robert Lightroot. NASA Associate Administrator; Dr. Roger Launius, Associate Director of Collections and Curatorial Affairs at the Smithsonian National Air and Space Museum; Dr. Harriett Jenkins, Former Assistant Administrator for Equal Opportunity Programs at NASA; Brenda Manuel, Associate Administrator for Diversity and Equal Opportunity at NASA; Suzanne Malveaux, CNN Correspondent and panel moderator; U.S. Representative Eddie Bernice Johnson, of Texas; Dr. Michael Eric Dyson, professor of sociology at Georgetown University; and Charles Bolden, NASA Administrator; pose for a picture following an event celebrating the 50th Anniversary of the Civil Rights Act of 1964 on Monday, June 23, 2014 in the James E. Webb Auditorium at NASA Headquarters in Washington, DC. The event highlighted the influence of the Civil Rights Act on NASA. Photo Credit: (NASA/Joel Kowsky)

Evaluation of cimi-shield knock-out bed bug eliminator against house fly (Musca domestica) adults

USDA-ARS?s Scientific Manuscript database

Cimi-Shield Knock-Out (CSKO) Bed Bug Eliminator is a green treatment labeled for use against bed bugs, carpet beetles, ants, roaches, fleas, ticks, silverfish, millipedes and centipedes. The active ingredient is soybean oil. If CSKO is formulated according to label instructions and sprayed directly ...

"When Do I Knock on the Hotel Room Door?": The MLA Conference Job Interview.

ERIC Educational Resources Information Center

Emmerson, Richard K.

1995-01-01

Offers advice to potential interviewees attending an MLA conference. Cautions the interviewee to arrive for the appointment early, but not to knock until the time of the appointment. Advises interviewees to answer questions briefly and to let the committee set the pace of the interview. (PA)

Evaluation of cimi-shield knock-out bed bug eliminator against house fly (Musca domestica) adults.

USDA-ARS?s Scientific Manuscript database

Cimi-Shield Knock-Out (CSKO) Bed Bug Eliminator is a green treatment labeled for use against bed bugs, carpet beetles, ants, roaches, fleas, ticks, silverfish, millipedes and centipedes. The active ingredient is soybean oil. If CSKO is formulated according to label instructions and sprayed directly ...

The Science of Hard Knocks

ERIC Educational Resources Information Center

Vance, Erik

2007-01-01

Head injuries in sports are nothing new, but in recent years, college athletes have reported a steady rise in concussions. Football players still get the most knocks to the head. Women have managed to keep up with, and often surpass, men in sports-related concussions in the last few years. In basketball, women reported 24 percent more concussions…

Performance of lignin derived compounds as octane boosters

DOE PAGES

Tian, Miao; McCormick, Robert L.; Ratcliff, Matthew A.; ...

2016-11-01

The performance of spark ignition engines is highly dependent on fuel anti-knock quality, which in turn is governed by autoignition chemistry. In this study, we explore this chemistry for various aromatic oxygenates (i.e., anisole, 4-methyl anisole, 4-propyl anisole, guaiacol, 4-methyl guaiacol, 4-ethyl guaiacol) that can be produced from lignin, a low value residual biomass stream that is generated in paper pulping and cellulosic ethanol plants. All compounds share the same benzene ring, but have distinct oxygen functionalities and degrees of alkylation. The objective of this study is to ascertain what the impact is of said side groups on anti-knock qualitymore » and, by proxy, on fuel economy in a modern Volvo T5 spark ignition engine. To better comprehend the variation in behavior amongst the fuels, further experiments have been conducted in a constant volume autoignition device. In conclusion, the results demonstrate that alkylation has a negligible impact on anti-knock quality, while the addition of functional oxygen groups manifests as a deterioration in anti-knock quality.« less

Monte Carlo analysis of the oxygen knock-on effects induced by synchrotron x-ray radiation in the B i2S r2CaC u2O8 +δ superconductor

NASA Astrophysics Data System (ADS)

Torsello, Daniele; Mino, Lorenzo; Bonino, Valentina; Agostino, Angelo; Operti, Lorenza; Borfecchia, Elisa; Vittone, Ettore; Lamberti, Carlo; Truccato, Marco

2018-01-01

We investigate the microscopic mechanism responsible for the change of macroscopic electrical properties of the B i2S r2CaC u2O8 +δ high-temperature superconductor induced by intense synchrotron hard x-ray beams. The possible effects of secondary electrons on the oxygen content via the knock-on interaction are studied by Monte Carlo simulations. The change in the oxygen content expected from the knock-on model is computed convoluting the fluence of photogenerated electrons in the material with the Seitz-Koehler cross section. This approach has been adopted to analyze several experimental irradiation sessions with increasing x-ray fluences. A close comparison between the expected variations in oxygen content and the experimental results allows determining the irradiation regime in which the knock-on mechanism can satisfactorily explain the observed changes. Finally, we estimate the threshold displacement energy of loosely bound oxygen atoms in this material Td=0 .15-0.01+0.025eV .

A novel luciferase knock-in reporter system for studying transcriptional regulation of the human Sox2 gene.

PubMed

Xiao, Dan; Zhang, Weifeng; Li, Yan; Liu, Kuan; Zhao, Junli; Sun, Xiaohong; Shan, Linlin; Mao, Qinwen; Xia, Haibin

2016-02-10

Sox2 is an important transcriptional factor that has multiple functions in stem cell maintenance and tumorigenesis. To investigate the transcriptional regulation of the Sox2 gene, a luciferase knock-in reporter system was established in HEK293 cells by placing the luciferase gene in the genome under the control of the Sox2 gene promoter using a transcription activator-like effector nuclease (TALEN)-mediated genome editing technique. PCR and Southern blot results confirmed the site-specific integration of a single copy of the exogenous luciferase gene into the genome. To prove the reliability and sensitivity of this novel luciferase knock-in system, a CRISPR/Cas transcription activation system for the Sox2 gene was constructed and applied to the knock-in system. The results indicated that luciferase activity was directly correlated with the activity of the Sox2 endogenous promoter. This novel system will be a useful tool to study the transcriptional regulation of Sox2, and has great potential in medical and industrial applications. Copyright © 2015 Elsevier B.V. All rights reserved.

Mild deficits in mice lacking pituitary adenylate cyclase-activating polypeptide receptor type 1 (PAC1) performing on memory tasks.

PubMed

Sauvage, M; Brabet, P; Holsboer, F; Bockaert, J; Steckler, T

2000-12-08

Pituitary adenylate cyclase-activating polypeptide (PACAP) and its receptor subtype 1 (PAC1) have been suggested to play a role in the modulation of learning and memory. However, behavioral evidence for altered mnemonic function due to altered PAC1 activity is missing. Therefore, the role of PAC1 in learning and memory was studied in mouse mutants lacking this receptor (PAC1 knock-out mice), tested in water maze two-choice spatial discrimination, one-trial contextual and cued fear conditioning, and multiple-session contextual discrimination. Water maze spatial discrimination was unaffected in PAC1 mutants, while a mild deficit was observed in multiple session contextual discrimination in PAC1 knock-out mice. Furthermore, PAC1 knock-out mice were able to learn the association between context and shock in one-trial contextual conditioning, but showed faster return to baseline than wild-type mice. Thus, the effects of PAC1 knock-out on modulating performance in these tasks were subtle and suggest that PAC1 only plays a limited role in learning and memory.

Als2 mRNA splicing variants detected in KO mice rescue severe motor dysfunction phenotype in Als2 knock-down zebrafish.

PubMed

Gros-Louis, Francois; Kriz, Jasna; Kabashi, Edor; McDearmid, Jonathan; Millecamps, Stéphanie; Urushitani, Makoto; Lin, Li; Dion, Patrick; Zhu, Qinzhang; Drapeau, Pierre; Julien, Jean-Pierre; Rouleau, Guy A

2008-09-01

Recessive ALS2 mutations are linked to three related but slightly different neurodegenerative disorders: amyotrophic lateral sclerosis, hereditary spastic paraplegia and primary lateral sclerosis. To investigate the function of the ALS2 encoded protein, we generated Als2 knock-out (KO) mice and zAls2 knock-down zebrafish. The Als2(-/-) mice lacking exon 2 and part of exon 3 developed mild signs of neurodegeneration compatible with axonal transport deficiency. In contrast, zAls2 knock-down zebrafish had severe developmental abnormalities, swimming deficits and motor neuron perturbation. We identified, by RT-PCR, northern and western blotting novel Als2 transcripts in mouse central nervous system. These Als2 transcripts were present in Als2 null mice as well as in wild-type littermates and some rescued the zebrafish phenotype. Thus, we speculate that the newly identified Als2 mRNA species prevent the Als2 KO mice from developing severe neurodegenerative disease and might also regulate the severity of the motor neurons phenotype observed in ALS2 patients.

MONOAMINE OXIDASE: From Genes to Behavior

PubMed Central

Shih, J. C.; Chen, K.; Ridd, M. J.

2010-01-01

Cloning of MAO (monoamine oxidase) A and B has demonstrated unequivocally that these enzymes are made up of different polypeptides, and our understanding of MAO structure, regulation, and function has been significantly advanced by studies using their cDNA. MAO A and B genes are located on the X-chromosome (Xp11.23) and comprise 15 exons with identical intron-exon organization, which suggests that they are derived from the same ancestral gene. MAO A and B knockout mice exhibit distinct differences in neurotransmitter metabolism and behavior. MAO A knock-out mice have elevated brain levels of serotonin, norephinephrine, and dopamine and manifest aggressive behavior similar to human males with a deletion of MAO A. In contrast, MAO B knock-out mice do not exhibit aggression and only levels of phenylethylamine are increased. Mice lacking MAO B are resistant to the Parkinsongenic neurotoxin, 1-methyl-4-phenyl-1,2,3,6-tetra-hydropyridine. Both MAO A and B knock-out mice show increased reactivity to stress. These knock-out mice are valuable models for investigating the role of monoamines in psychoses and neurodegenerative and stress-related disorders. PMID:10202537

Clinical practice: new challenges for the advanced practice nurse.

PubMed

Bartel, J C; Buturusis, B

2000-12-01

This report describes the challenges for advanced practice nurses (APNs) relative to supply and demand issues. The article also includes opportunities with the Balanced Budget Act, physician acceptance of Advanced Practice Nurses, and expanding practice opportunities. The challenges include the nursing shortage (both in nursing students and faculty), the aging of the nursing workforce, and a lag in nursing salaries; increased demand for nursing based on aging baby boomers, increasing patient acuity and technology, and new arenas for practice. The Balanced Budget Act of 1997 provided new opportunities for advanced practice nurses, including enhanced autonomy to provide services and bill independently of physicians. With these changes come new opportunities for advanced practice nurse entrepreneurs in the areas of independent practice, including opportunities to positively impact the health of families and communities in alignment with the Federal government's vision for "Healthy People 2010." As physician acceptance of advanced practice nurses continues to grow and in light of the changes in medical practice and education (residency reduction), opportunities to expand collaborative practice arrangements also exist. APNs are best suited to make the most of these changes. One example of an opportunity for independent practice, a Community Wellness Center, is developed as an entrepreneurial venture benefiting both the APN and the health of a community. Who better than registered nurses (RNs), especially those practicing at the advanced level, can ensure that these opportunities and challenges are addressed in an ethical manner and focused on the needs and health of the community?

Global Climate Change - U.S. Economic and National Security Opportunity

DTIC Science & Technology

2009-03-20

The most recent findings of the Intergovernmental Panel on Climate Change (IPCC) state that the current trajectory of greenhouse gas (GHG) emissions...challenges and opportunities for the United States as they balance national security and economic interests. The effects of climate change could act as a...are various opportunities associated with climate change including opening arctic navigational channels and the vast oil and natural gas resources

[Expression of matrix metalloproteinase-19 in the human cornea. Wound healing in the MMP-19 knock-out mouse model].

PubMed

Treumer, F; Flöhr, C; Klettner, A; Nölle, B; Roider, J

2010-07-01

At present there are no data in the literature on the expression of matrix metalloprotein-19 in the human cornea. The aim of this study was to analyze the expression of matrix metalloproteinase-19 in the human cornea and to investigate its potential role in corneal wound healing using a MMP-19 knock-out mouse model. A method with Western blotting and immunohistological staining for MMP-19 was performed using paraffin embedded human corneas. Excimer laser keratectomy was performed in wild type (wt) and MMP-19 knock-out (ko) mice and the rate of re-epithelialization was analyzed after 8 h and 18 h. MMP-19 was strongly expressed in the human corneal epithelium mainly in the basal cell layer. MMP-19 was not expressed in the corneal stroma. In the mouse model the size of the corneal lesion after 8 h was 83% (wt) and 89.9% (ko) of the initial area (p=0.09). After 18 h the lesion was 17% (wt) and 13.3% (ko) of the initial area (p=0.01). Laminin-5 was expressed in the migrating epithelial cells with no differences between wild type and knock-out mouse. MMP-19 showed a strong expression in the basal cells of the human corneal epithelium. Corneal re-epithelialization was slightly faster in the MMP-19 knock-out mouse. No differences in the expression of laminin-5 could be detected.

Anti-knock quality of sugar derived levulinic esters and cyclic ethers

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tian, Miao; McCormick, Robert L.; Luecke, Jon

Here, the objective of this paper is to investigate the anti-knock quality of sugar-derived levulinic esters (methyl levulinate (ML) and ethyl levulinate (EL)) and cyclic ethers (furfuryl ethyl ether (FEE) and ethyl tetrahydrofurfuryl ether (ETE)). To this end, combustion experiments were carried out in both an engine and a constant volume autoignition device. The results from both apparati demonstrate that ML, EL and FEE have superior anti-knock quality than the reference Euro95 gasoline. ETE, conversely, performed markedly worse than the reference fuel on both setups and might therefore be a more appropriate fuel for compression ignition engines. The main reasonmore » of the distinctions in anti-knock quality can be found in the molecular structure of the neat biofuels. ML and EL are levulinic esters, with a ketone (C=O) functionality and an ester (C(=O)-O) group on the carbon chain. They can readily produce stable intermediates during the auto-ignition process, thereby slowing down the overall reaction rate. The unsaturated cyclic ether (FEE) has very strong ring C-H bonds. However, the saturated cyclic ether (ETE) has weak ring C-H bonds, which facilitate more readily ring opening reactions. Long side chains on the cyclic ethers further accelerate the reaction rate. Importantly for future research, our results suggest that IQT and engine experiments are interchangeable setups with respect to qualitative anti-knock quality evaluation of novel compounds.« less

The Effects of Engine Speed and Mixture Temperature on the Knocking Characteristics of Several Fuels

NASA Technical Reports Server (NTRS)

Lee, Dana W

1940-01-01

Six 100-octane and two 87-octane aviation engine fuels were tested in a modified C.F.R. variable-compression engine at 1,500, 2,000 and 2,500 rpm. The mixture temperature was raised from 50 to 300 F in approximately 50 degree steps and, at each temperature, the compression ratio was adjusted to give incipient knock as shown by a cathode ray indicator. The results are presented in tabular form. The results are analyzed on the assumption that the conditions which determine whether a given fuel will knock are the maximum values of density and temperature reached by the burning gases. A maximum permissible density factor, proportional to the maximum density of the burning gases just prior to incipient knock, and the temperature of the burning gases at that time were computed for each of the test conditions. Values of the density factors were plotted against the corresponding end-gas temperatures for the three engine speeds and also against engine speed for several and end-gas temperatures. The maximum permissible density factor varied only slightly with engine speed but decreased rapidly with an increase in the end-gas temperature. The effect of changing the mixture temperature was different for fuels of different types. The results emphasize the desirability of determining the anti knock values of fuels over a wide range of engine and intake-air conditions rather that at a single set of conditions.

CRISPR/Cas9 Mediated GFP Knock-in at the MAP1LC3B Locus in 293FT Cells Is Better for Bona Fide Monitoring Cellular Autophagy.

PubMed

Wu, Zhiqiang; Zhao, Jinlin; Qiu, Minghan; Mi, Zeyun; Meng, Maobin; Guo, Yu; Wang, Hui; Yuan, Zhiyong

2018-04-19

Accurately identifying and quantifying cellular autophagy is very important as the significance of autophagy in physiological and pathological processes becomes increasingly evident. Ectopically expressed fluorescent-tagged microtubule-associated protein light chain 3B (MAP1LC3B, LC3) is the most widely used reporter for monitoring autophagy activity thus far. However, this approach ignores the influence of constitutively overexpressed LC3 on autophagy itself and autophagy-related processes and its accuracy in indicating autophagy is questionable. Here, we generated a knock-in GFP-LC3 reporter via the CRISPR/Cas9 system in 293FT cells to add GFP to the N-terminal of and in frame with endogenous LC3. We proved that this knock-in GFP-LC3 was expressed at biological level driven by the endogenous transcriptional regulatory elements as the wild type alleles. Compared with the ectopically expressed GFP-LC3, the endogenous knock-in reporter exhibited much higher sensitivity and signal-to-noise ratio of GFP-LC3 puncta upon the induction or inhibition of autophagy at certain step for monitoring autophagy activity. Thus, according to the previous reported concerning and the results presented here, we suggest that this knock-in GFP-LC3 reporter is better for bona fide monitoring cellular autophagy and should be employed for further study of autophagy in vitro and in vivo. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A Simple Retroelement Based Knock-Down System in Dictyostelium: Further Insights into RNA Interference Mechanisms.

PubMed

Friedrich, Michael; Meier, Doreen; Schuster, Isabelle; Nellen, Wolfgang

2015-01-01

We have previously shown that the most abundant Dictyostelium discoideum retroelement DIRS-1 is suppressed by RNAi mechanisms. Here we provide evidence that both inverted terminal repeats have strong promoter activity and that bidirectional expression apparently generates a substrate for Dicer. A cassette containing the inverted terminal repeats and a fragment of a gene of interest was sufficient to activate the RNAi response, resulting in the generation of ~21 nt siRNAs, a reduction of mRNA and protein expression of the respective endogene. Surprisingly, no transitivity was observed on the endogene. This was in contrast to previous observations, where endogenous siRNAs caused spreading on an artificial transgene. Knock-down was successful on seven target genes that we examined. In three cases a phenotypic analysis proved the efficiency of the approach. One of the target genes was apparently essential because no knock-out could be obtained; the RNAi mediated knock-down, however, resulted in a very slow growing culture indicating a still viable reduction of gene expression. ADVANTAGES OF THE DIRS-1–RNAI SYSTEM: The knock-down system required a short DNA fragment (~400 bp) of the target gene as an initial trigger. Further siRNAs were generated by RdRPs since we have shown some siRNAs with a 5'-triphosphate group. Extrachromosomal vectors facilitate the procedure and allowed for molecular and phenotypic analysis within one week. The system provides an efficient and rapid method to reduce protein levels including those of essential genes.

Highly efficient generation of knock-in transgenic medaka by CRISPR/Cas9-mediated genome engineering.

PubMed

Watakabe, Ikuko; Hashimoto, Hisashi; Kimura, Yukiko; Yokoi, Saori; Naruse, Kiyoshi; Higashijima, Shin-Ichi

2018-01-01

Medaka ( Oryzias latipes ) is a popular animal model used in vertebrate genetic analysis. Recently, an efficient (~ 30%) knock-in system via non-homologous end joining (NHEJ) was established in zebrafish using the CRISPR/Cas9 system. If the same technique were applicable in medaka, it would greatly expand the usefulness of this model organism. The question of the applicability of CRISPR/Cas9 in medaka, however, has yet to be addressed. We report the highly efficient generation of knock-in transgenic medaka via non-homologous end joining (NHEJ). Donor plasmid containing a heat-shock promoter and a reporter gene was co-injected with a short guide RNA (sgRNA) targeted for genome digestion, an sgRNA targeted for donor plasmid digestion, and Cas9 mRNA. Broad transgene expression in the expression domain of a target gene was observed in approximately 25% of injected embryos. By raising these animals, we established stable knock-in transgenic fish with several different constructs for five genetic loci, obtaining transgenic founders at efficiencies of > 50% for all five loci. Further, we show that the method is useful for obtaining mutant alleles. In the experiments where transgene integrations were targeted between the transcription start site and the initiation methionine, the resultant transgenic fish became mutant alleles. With its simplicity, design flexibility, and high efficiency, we propose that CRISPR/Cas9-mediated knock-in via NHEJ will become a standard method for the generation of transgenic and mutant medaka.

Knock-in strategy at 3'-end of Crx gene by CRISPR/Cas9 system shows the gene expression profiles during human photoreceptor differentiation.

PubMed

Homma, Kohei; Usui, Sumiko; Kaneda, Makoto

2017-03-01

Fluorescent reporter gene knock-in induced pluripotent stem cell (iPSC) lines have been used to evaluate the efficiency of differentiation into specific cell lineages. Here, we report a knock-in strategy for the generation of human iPSC reporter lines in which a 2A peptide sequence and a red fluorescent protein (E2-Crimson) gene were inserted at the termination codon of the cone-rod homeobox (Crx) gene, a photoreceptor-specific transcriptional factor gene. The knock-in iPSC lines were differentiated into fluorescence-expressing cells in 3D retinal differentiation culture, and the fluorescent cells also expressed Crx specifically in the nucleus. We found that the fluorescence intensity was positively correlated with the expression levels of Crx mRNA and that fluorescent cells expressed rod photoreceptor-specific genes in the later stage of differentiation. Finally, we treated the fluorescent cells with DAPT, a Notch inhibitor, and found that DAPT-enhanced retinal differentiation was associated with up-regulation of Crx, Otx2 and NeuroD1, and down-regulation of Hes5 and Ngn2. These suggest that this knock-in strategy at the 3'-end of the target gene, combined with the 2A peptide linked to fluorescent proteins, offers a useful tool for labeling specific cell lineages or monitoring expression of any marker genes without affecting the function of the target gene. © 2017 Molecular Biology Society of Japan and John Wiley & Sons Australia, Ltd.

Anti-knock quality of sugar derived levulinic esters and cyclic ethers

DOE PAGES

Tian, Miao; McCormick, Robert L.; Luecke, Jon; ...

2017-04-22

Here, the objective of this paper is to investigate the anti-knock quality of sugar-derived levulinic esters (methyl levulinate (ML) and ethyl levulinate (EL)) and cyclic ethers (furfuryl ethyl ether (FEE) and ethyl tetrahydrofurfuryl ether (ETE)). To this end, combustion experiments were carried out in both an engine and a constant volume autoignition device. The results from both apparati demonstrate that ML, EL and FEE have superior anti-knock quality than the reference Euro95 gasoline. ETE, conversely, performed markedly worse than the reference fuel on both setups and might therefore be a more appropriate fuel for compression ignition engines. The main reasonmore » of the distinctions in anti-knock quality can be found in the molecular structure of the neat biofuels. ML and EL are levulinic esters, with a ketone (C=O) functionality and an ester (C(=O)-O) group on the carbon chain. They can readily produce stable intermediates during the auto-ignition process, thereby slowing down the overall reaction rate. The unsaturated cyclic ether (FEE) has very strong ring C-H bonds. However, the saturated cyclic ether (ETE) has weak ring C-H bonds, which facilitate more readily ring opening reactions. Long side chains on the cyclic ethers further accelerate the reaction rate. Importantly for future research, our results suggest that IQT and engine experiments are interchangeable setups with respect to qualitative anti-knock quality evaluation of novel compounds.« less

Cholecystokinin levels in prohormone convertase 2 knock-out mouse brain regions reveal a complex phenotype of region-specific alterations.

PubMed

Beinfeld, Margery C; Blum, Alissa; Vishnuvardhan, Daesety; Fanous, Sanya; Marchand, James E

2005-11-18

Prohormone convertase 2 is widely co-localized with cholecystokinin in rodent brain. To examine its role in cholecystokinin processing, cholecystokinin levels were measured in dissected brain regions from prohormone convertase 2 knock-out mice. Cholecystokinin levels were lower in hippocampus, septum, thalamus, mesencephalon, and pons in knock-out mice than wild-type mice. In cerebral cortex, cortex-related structures and olfactory bulb, cholecystokinin levels were higher than wild type. Female mice were more affected by the loss of prohormone convertase 2 than male mice. The decrease in cholecystokinin levels in these brain regions shows that prohormone convertase 2 is important for cholecystokinin processing. Quantitative polymerase chain reaction measurements were performed to examine the relationship between peptide levels and cholecystokinin and enzyme expression. They revealed that cholecystokinin and prohormone convertase 1 mRNA levels in cerebral cortex and olfactory bulb were actually lower in knock-out than wild type, whereas their expression in other brain regions of knock-out mouse brain was the same as wild type. Female mice frequently had higher expression of cholecystokinin and prohormone convertase 1, 2, and 5 mRNA than male mice. The loss of prohormone convertase 2 alters CCK processing in specific brain regions. This loss also appears to trigger compensatory mechanisms in cerebral cortex and olfactory bulb that produce elevated levels of cholecystokinin but do not involve increased expression of cholecystokinin, prohormone convertase 1 or 5 mRNA.

Every Student Succeeds Act (ESSA): New Legislation Creates Opportunities for Parent Advocates

ERIC Educational Resources Information Center

Welch, Carolyn E.

2016-01-01

The passage of the Every Student Succeeds Act (ESSA) in December 2015 is an exciting development for parents, teachers, school leaders, and others who believe U.S. schools should meet the needs of high-ability students. The ESSA revised and reauthorized the Elementary and Secondary Education Act of 1965 (ESEA), previously known as the No Child…

29 CFR 1621.4 - Effect of opinions and interpretations of the Commission.

Code of Federal Regulations, 2010 CFR

2010-07-01

.... 1621.4 Section 1621.4 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-THE EQUAL PAY ACT § 1621.4 Effect of opinions and interpretations of the Commission. (a) Section 10 of the Portal to Portal Act of 1947, 29 U.S.C. 255, which applies to the Equal Pay Act of 1963...

29 CFR 37.3 - How does this part affect a recipient's other obligations?

Code of Federal Regulations, 2010 CFR

2010-07-01

... Vietnam Era Veterans' Readjustment Assistance Act of 1974, as amended (38 U.S.C. 4212); (4) The Equal Pay... EQUAL OPPORTUNITY PROVISIONS OF THE WORKFORCE INVESTMENT ACT OF 1998 (WIA) General Provisions § 37.3 How... regulations implementing Title VI of the Civil Rights Act of 1964, as amended (Title VI), and with Subparts A...

The Perceived Impact of Education Policy on the Inclusion of Students with Disabilities in Two Contrasting Schools

ERIC Educational Resources Information Center

Foster, Megan Holmwood

2013-01-01

It has been nearly four decades since the passage of the Education for All Handicapped Children Act (1975), yet some of the most fundamental issues still remain. Reauthorized in 1997, and again in 2004, as the Individuals with Disabilities Act (IDEA), this act requires all children with disabilities be offered educational opportunities in the…

Adult Education and Family Literacy Act. Report to Congress on State Performance. Program Year 2003-2004

ERIC Educational Resources Information Center

Office of Vocational and Adult Education, US Department of Education, 2006

2006-01-01

The state-administered grant program authorized under the Adult Education and Family Literacy Act (AEFLA), enacted as Title II of the Workforce Investment Act (WIA) of 1998, is the major source of federal support for adult basic and literacy education programs. The purpose of the program is to provide educational opportunities to adults sixteen…

78 FR 80415 - To Take Certain Actions Under the African Growth and Opportunity Act and for Other Purposes

Federal Register 2010, 2011, 2012, 2013, 2014

2013-12-31

..., 1985, the United States and Israel entered into the Agreement on the Establishment of a Free Trade Area between the Government of the United States of America and the Government of Israel (USIFTA), which the Congress approved in the United States-Israel Free Trade Area Implementation Act of 1985 (the ``USIFTA Act...

75 FR 81077 - To Take Certain Actions Under the African Growth and Opportunity Act, and for Other Purposes

Federal Register 2010, 2011, 2012, 2013, 2014

2010-12-27

... States and Israel entered into the Agreement on the Establishment of a Free Trade Area between the Government of the United States of America and the Government of Israel (the ``USIFTA''), which the Congress approved in the United States-Israel Free Trade Area Implementation Act of 1985 (the ``USIFTA Act'') (19 U...

Healthy, hunger-free kids act of 2010: an opportunity for school nurses to make a difference.

PubMed

Prokop, Jessica L; Galon, Patricia

2011-12-01

Implementation of the Healthy Hunger-Free Kids Act of 2010 will provide an opportunity for school nurses to intervene in the serious childhood obesity problem in the United States. Major changes in the management of the National School Lunch Program (NSLP) will likely challenge schools yet may provide the impetus for a collaborative effort by the interdisciplinary school staff and parents to make a real difference for children. School nurses must use this occasion for a change to promote healthy eating habits and increased physical activity levels for students to carry into adulthood.

A Healthy New World: Emergent Opportunities for Clinical Psychologists in the Patient Protection and Affordable Care Act

PubMed Central

Beidas, Rinad S.; Manderscheid, Ronald W.

2014-01-01

The Patient Protection and Affordable Care Act (ACA) is radically transforming the health and mental health care landscape. Emergent opportunities exist for clinical psychologists to redefine their role in healthcare. We reflect on the Chor and colleagues article (this issue) elucidating key issues for psychologists, and present additional recommendations for consideration. Specifically, we highlight three points: (1) moving beyond just training and hoping; (2) recovery, not just symptom reduction; and (3) it’s a healthy new world. Under each of these points, we suggest tactics for how to achieve these goals. PMID:24954983

29 CFR 1626.20 - Procedure for requesting an opinion letter.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 1626.20 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN EMPLOYMENT ACT § 1626.20 Procedure for requesting an opinion letter. (a) A request for an opinion letter should be submitted in writing to the Chairman, Equal Employment Opportunity...

29 CFR 1626.20 - Procedure for requesting an opinion letter.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 1626.20 Labor Regulations Relating to Labor (Continued) EQUAL EMPLOYMENT OPPORTUNITY COMMISSION PROCEDURES-AGE DISCRIMINATION IN EMPLOYMENT ACT § 1626.20 Procedure for requesting an opinion letter. (a) A request for an opinion letter should be submitted in writing to the Chairman, Equal Employment Opportunity...