Science.gov

Sample records for live cold-adapted flumist

  1. Influenza vaccination for the pediatric patient: a focus on the new intranasal, cold-adapted, live attenuated vaccine.

    PubMed

    Ellis, Jennifer M; Reilly, Joan C; Salazar, Juan C

    2004-04-01

    FluMist is the first live attenuated, cold-adapted intranasal influenza vaccine (LAIV) approved for the prevention of influenza A and B. Clinical trials have shown that annual vaccination with LAIV is effective for the prevention of influenza. LAIV appears well tolerated in healthy patients 5-49 years of age. The most common adverse events are abdominal pain, chills, cough, diarrhea, headache, irritability, lethargy, muscle aches, otitis media, rhinitis, sinusitis, sore throat, and vomiting. FluMist has a novel intranasal route of administration that allows for influenza prevention without a painful intramuscular injection. Barriers preventing acceptance of LAIV include defining the appropriate patient population, cost, and insurance coverage.

  2. Demystifying FluMist, a new intranasal, live influenza vaccine.

    PubMed

    Mossad, Sherif B

    2003-09-01

    FluMist--a cold-adapted, live-attenuated, trivalent, intranasal influenza virus vaccine approved by the US Food and Drug Administration on June 17, 2003--has been shown to be safe and effective, but its role in the general prevention of influenza is yet to be defined. Intranasal administration is expected to be more acceptable than parenteral, particularly in children, but the potential for the shedding of live virus may pose a risk to anyone with a compromised immune system. PMID:14518575

  3. Live cold-adapted influenza A vaccine produced in Vero cell line.

    PubMed

    Romanova, Julia; Katinger, Dietmar; Ferko, Boris; Vcelar, Brigitta; Sereinig, Sabine; Kuznetsov, Oleg; Stukova, Marina; Erofeeva, Marjana; Kiselev, Oleg; Katinger, Hermann; Egorov, Andrej

    2004-07-01

    The African green monkey kidney (Vero) cell line was used as a substrate for the development of a live cold-adapted (ca) reassortant influenza vaccine. For that purpose, a new master strain was generated by an adaptation of the wild type (wt) A/Singapore/1/57 virus to growth at 25 degrees C in a Vero cell line. The resulting cold-adapted (ca) muster strain A/Singapore/1/57ca showed temperature sensitive (ts) phenotype and was attenuated in animal models and protective in the challenge experiments in ferrets. Two vaccine candidates of influenza A(H1N1) and A(H3N2) subtypes (6/2 reassortants) inheriting six genes coding internal proteins from the new master strain and the surface antigens hemagglutinin (HA) and neuraminidase (NA) from the epidemic viruses were obtained by a standard method of genetic reassortment. All steps of the vaccine preparation were done exclusively in Vero cells, including the isolation of the epidemic viruses. Both vaccine strains were used for immunization of young adult volunteers in a limited clinical trial and appeared to be safe, well tolerated and immunogenic after intranasal administration. PMID:15163508

  4. Enhancement of the safety of live influenza vaccine by attenuating mutations from cold-adapted hemagglutinin.

    PubMed

    Lee, Yoon Jae; Jang, Yo Han; Kim, Paul; Lee, Yun Ha; Lee, Young Jae; Byun, Young Ho; Lee, Kwang-Hee; Kim, Kyusik; Seong, Baik Lin

    2016-04-01

    In our previous study, X-31ca-based H5N1 LAIVs, in particular, became more virulent in mice than the X-31ca MDV, possibly by the introduction of the surface antigens of highly pathogenic H5N1 influenza virus, implying that additional attenuation is needed in this cases to increase the safety level of the vaccine. In this report we suggest an approach to further increase the safety of LAIV through additional cold-adapted mutations in the hemagglutinin. The cold-adaptation of X-31 virus resulted in four amino acid mutations in the HA. We generated a panel of 7:1 reassortant viruses each carrying the hemagglutinins with individual single amino acid mutations. We examined their phenotypes and found a major attenuating mutation, N81K. This attenuation marker conferred additional temperature-sensitive and attenuation phenotype to the LAIV. Our data indicate that the cold-adapted mutation in the HA confers additional attenuation to the LAIV strain, without compromising its productivity and immune response.

  5. The cold adapted and temperature sensitive influenza A/Ann Arbor/6/60 virus, the master donor virus for live attenuated influenza vaccines, has multiple defects in replication at the restrictive temperature

    SciTech Connect

    Chan, Winnie; Zhou, Helen; Kemble, George; Jin Hong

    2008-10-25

    We have previously determined that the temperature sensitive (ts) and attenuated (att) phenotypes of the cold adapted influenza A/Ann Arbor/6/60 strain (MDV-A), the master donor virus for the live attenuated influenza A vaccines (FluMist), are specified by the five amino acids in the PB1, PB2 and NP gene segments. To understand how these loci control the ts phenotype of MDV-A, replication of MDV-A at the non-permissive temperature (39 deg. C) was compared with recombinant wild-type A/Ann Arbor/6/60 (rWt). The mRNA and protein synthesis of MDV-A in the infected MDCK cells were not significantly reduced at 39 deg. C during a single-step replication, however, vRNA synthesis was reduced and the nuclear-cytoplasmic export of viral RNP (vRNP) was blocked. In addition, the virions released from MDV-A infected cells at 39 deg. C exhibited irregular morphology and had a greatly reduced amount of the M1 protein incorporated. The reduced M1 protein incorporation and vRNP export blockage correlated well with the virus ts phenotype because these defects could be partially alleviated by removing the three ts loci from the PB1 gene. The virions and vRNPs isolated from the MDV-A infected cells contained a higher level of heat shock protein 70 (Hsp70) than those of rWt, however, whether Hsp70 is involved in thermal inhibition of MDV-A replication remains to be determined. Our studies demonstrate that restrictive replication of MDV-A at the non-permissive temperature occurs in multiple steps of the virus replication cycle.

  6. Efficacy and effectiveness of live attenuated influenza vaccine in school-age children.

    PubMed

    Coelingh, Kathleen; Olajide, Ifedapo Rosemary; MacDonald, Peter; Yogev, Ram

    2015-01-01

    Evidence of high efficacy of live attenuated influenza vaccine (LAIV) from randomized controlled trials is strong for children 2-6 years of age, but fewer data exist for older school-age children. We reviewed the published data on efficacy and effectiveness of LAIV in children ≥5 years. QUOSA (Elsevier database) was searched for articles published from January 1990 to June 2014 that included 'FluMist', 'LAIV', 'CAIV', 'cold adapted influenza vaccine', 'live attenuated influenza vaccine', 'live attenuated cold adapted' or 'flu mist'. Studies evaluated included randomized controlled trials, effectiveness and indirect protection studies. This review demonstrates that LAIV has considerable efficacy and effectiveness in school-age children.

  7. Serological evaluation of an influenza A virus cold-adapted reassortant live vaccine, CR-37 (H1N1), in Japanese adult volunteers.

    PubMed

    Yamane, N; Nakamura, Y; Yuki, M; Odagiri, T; Ishida, N

    1984-04-01

    A cold-adapted influenza A virus, CR-37 (H1N1), derived from genetic reassortment between A/Ann Arbor/6/60 (H2N2) cold-adapted variant virus and A/California/10/78 (H1N1) wild-type virus, was tested in Japanese adult volunteer. The CR-37 live virus preparation induced only low-grade clinical reactions in volunteers for the first 3-4 days after inoculation. Two vaccinees who did not show any antibody changes became febrile (over 38.0 degrees C). Skin tests using the vaccine preparation and uninfected allantoic fluid were performed, and indicated that one of these two vaccines was positive for the CR-37 vaccine preparation. A high proportion of the vaccinees whose sera had a haemagglutination-inhibition (HI) antibody titre against the vaccine strain of less than or equal to 64 before inoculation, seroconverted in both HI and neuraminidase-inhibition (NAI) antibody titrations, and only a few seroconverted in the titration of antibody against type-specific internal antigens. The serological examinations against heterotypic H1N1 variants indicated that the cold-adapted live influenza virus vaccine could induce a broad spectrum of HI antibody reactivity and immunity of long duration.

  8. Influenza virus vaccine live intranasal--MedImmune vaccines: CAIV-T, influenza vaccine live intranasal.

    PubMed

    2003-01-01

    MedImmune Vaccines (formerly Aviron) has developed a cold-adapted live influenza virus vaccine [FluMist] that can be administered by nasal spray. FluMist is the first live virus influenza vaccine and also the first nasally administered vaccine to be marketed in the US. The vaccine will be formulated to contain live attenuated (att) influenza virus reassortants of the strains recommended by the US Public Health Service for each 'flu season. The vaccine is termed cold-adapted (ca) because the virus has been adapted to replicate efficiently at 25 degrees C in the nasal passages, which are below normal body temperature. The strains used in the seasonal vaccine will also be made temperature sensitive (ts) so that their replication is restricted at 37 degrees C (Type B strains) and 39 degrees C (Type A strains). The combined effect of the antigenic properties and the att, ca and ts phenotypes of the influenza strains contained in the vaccine enables the viruses to replicate in the nasopharynx to produce protective immunity. The original formulation of FluMist requires freezer storage throughout distribution. Because many international markets do not have distribution channels well suited to the sale of frozen vaccines, Wyeth and MedImmune are collaborating to develop a second generation, refrigerator-stable, liquid trivalent cold-adapted influenza vaccine (CAIV-T), which is in phase III trials. Initially, the frozen formulation will only be available in the US. For the 2003-2004 season, FluMist will contain A/New Caledonia/20/99 (H1N1), A/Panama/2007/99 (H3N2) (A/Moscow/10/99-like) and B/Hong Kong/330/2001. Aviron was acquired by MedImmune on 15 January 2002. Aviron is now a wholly-owned subsidiary of MedImmune and is called MedImmune Vaccines. Aviron acquired FluMist in March 1995 through a Co-operative Research and Development Agreement (CRADA) with the US NIAID, and a licensing agreement with the University of Michigan, Ann Arbor, USA. In June 2000, the CRADA was

  9. Influenza virus vaccine live intranasal--MedImmune vaccines: CAIV-T, influenza vaccine live intranasal.

    PubMed

    2003-01-01

    MedImmune Vaccines (formerly Aviron) has developed a cold-adapted live influenza virus vaccine [FluMist] that can be administered by nasal spray. FluMist is the first live virus influenza vaccine and also the first nasally administered vaccine to be marketed in the US. The vaccine will be formulated to contain live attenuated (att) influenza virus reassortants of the strains recommended by the US Public Health Service for each 'flu season. The vaccine is termed cold-adapted (ca) because the virus has been adapted to replicate efficiently at 25 degrees C in the nasal passages, which are below normal body temperature. The strains used in the seasonal vaccine will also be made temperature sensitive (ts) so that their replication is restricted at 37 degrees C (Type B strains) and 39 degrees C (Type A strains). The combined effect of the antigenic properties and the att, ca and ts phenotypes of the influenza strains contained in the vaccine enables the viruses to replicate in the nasopharynx to produce protective immunity. The original formulation of FluMist requires freezer storage throughout distribution. Because many international markets do not have distribution channels well suited to the sale of frozen vaccines, Wyeth and MedImmune are collaborating to develop a second generation, refrigerator-stable, liquid trivalent cold-adapted influenza vaccine (CAIV-T), which is in phase III trials. Initially, the frozen formulation will only be available in the US. For the 2003-2004 season, FluMist will contain A/New Caledonia/20/99 (H1N1), A/Panama/2007/99 (H3N2) (A/Moscow/10/99-like) and B/Hong Kong/330/2001. Aviron was acquired by MedImmune on 15 January 2002. Aviron is now a wholly-owned subsidiary of MedImmune and is called MedImmune Vaccines. Aviron acquired FluMist in March 1995 through a Co-operative Research and Development Agreement (CRADA) with the US NIAID, and a licensing agreement with the University of Michigan, Ann Arbor, USA. In June 2000, the CRADA was

  10. Cold-adapted pandemic 2009 H1N1 influenza virus live vaccine elicits cross-reactive immune responses against seasonal and H5 influenza A viruses.

    PubMed

    Jang, Yo Han; Byun, Young Ho; Lee, Yoon Jae; Lee, Yun Ha; Lee, Kwang-Hee; Seong, Baik Lin

    2012-05-01

    The rapid transmission of the pandemic 2009 H1N1 influenza virus (pH1N1) among humans has raised the concern of a potential emergence of reassortment between pH1N1 and highly pathogenic influenza strains, especially the avian H5N1 influenza virus. Here, we report that the cold-adapted pH1N1 live attenuated vaccine (CApH1N1) elicits cross-reactive immunity to seasonal and H5 influenza A viruses in the mouse model. Immunization with CApH1N1 induced both systemic and mucosal antibodies with broad reactivity to seasonal and H5 strains, including HAPI H5N1 and the avian H5N2 virus, providing complete protection against heterologous and heterosubtypic lethal challenges. Our results not only accentuate the merit of using live attenuated influenza virus vaccines in view of cross-reactivity but also represent the potential of CApH1N1 live vaccine for mitigating the clinical severity of infections that arise from reassortments between pH1N1 and highly pathogenic H5 subtype viruses.

  11. Human whole body cold adaptation

    PubMed Central

    Daanen, Hein A.M.; Van Marken Lichtenbelt, Wouter D.

    2016-01-01

    ABSTRACT Reviews on whole body human cold adaptation generally do not distinguish between population studies and dedicated acclimation studies, leading to confusing results. Population studies show that indigenous black Africans have reduced shivering thermogenesis in the cold and poor cold induced vasodilation in fingers and toes compared to Caucasians and Inuit. About 40,000 y after humans left Africa, natives in cold terrestrial areas seems to have developed not only behavioral adaptations, but also physiological adaptations to cold. Dedicated studies show that repeated whole body exposure of individual volunteers, mainly Caucasians, to severe cold results in reduced cold sensation but no major physiological changes. Repeated cold water immersion seems to slightly reduce metabolic heat production, while repeated exposure to milder cold conditions shows some increase in metabolic heat production, in particular non-shivering thermogenesis. In conclusion, human cold adaptation in the form of increased metabolism and insulation seems to have occurred during recent evolution in populations, but cannot be developed during a lifetime in cold conditions as encountered in temperate and arctic regions. Therefore, we mainly depend on our behavioral skills to live in and survive the cold. PMID:27227100

  12. Human whole body cold adaptation.

    PubMed

    Daanen, Hein A M; Van Marken Lichtenbelt, Wouter D

    2016-01-01

    Reviews on whole body human cold adaptation generally do not distinguish between population studies and dedicated acclimation studies, leading to confusing results. Population studies show that indigenous black Africans have reduced shivering thermogenesis in the cold and poor cold induced vasodilation in fingers and toes compared to Caucasians and Inuit. About 40,000 y after humans left Africa, natives in cold terrestrial areas seems to have developed not only behavioral adaptations, but also physiological adaptations to cold. Dedicated studies show that repeated whole body exposure of individual volunteers, mainly Caucasians, to severe cold results in reduced cold sensation but no major physiological changes. Repeated cold water immersion seems to slightly reduce metabolic heat production, while repeated exposure to milder cold conditions shows some increase in metabolic heat production, in particular non-shivering thermogenesis. In conclusion, human cold adaptation in the form of increased metabolism and insulation seems to have occurred during recent evolution in populations, but cannot be developed during a lifetime in cold conditions as encountered in temperate and arctic regions. Therefore, we mainly depend on our behavioral skills to live in and survive the cold.

  13. Human whole body cold adaptation.

    PubMed

    Daanen, Hein A M; Van Marken Lichtenbelt, Wouter D

    2016-01-01

    Reviews on whole body human cold adaptation generally do not distinguish between population studies and dedicated acclimation studies, leading to confusing results. Population studies show that indigenous black Africans have reduced shivering thermogenesis in the cold and poor cold induced vasodilation in fingers and toes compared to Caucasians and Inuit. About 40,000 y after humans left Africa, natives in cold terrestrial areas seems to have developed not only behavioral adaptations, but also physiological adaptations to cold. Dedicated studies show that repeated whole body exposure of individual volunteers, mainly Caucasians, to severe cold results in reduced cold sensation but no major physiological changes. Repeated cold water immersion seems to slightly reduce metabolic heat production, while repeated exposure to milder cold conditions shows some increase in metabolic heat production, in particular non-shivering thermogenesis. In conclusion, human cold adaptation in the form of increased metabolism and insulation seems to have occurred during recent evolution in populations, but cannot be developed during a lifetime in cold conditions as encountered in temperate and arctic regions. Therefore, we mainly depend on our behavioral skills to live in and survive the cold. PMID:27227100

  14. A live attenuated cold-adapted influenza A H7N3 virus vaccine provides protection against homologous and heterologous H7 viruses in mice and ferrets

    SciTech Connect

    Joseph, Tomy; McAuliffe, Josephine; Lu, Bin; Vogel, Leatrice; Swayne, David; Jin, Hong; Kemble, George; Subbarao, Kanta

    2008-08-15

    The appearance of human infections caused by avian influenza A H7 subtype viruses underscores their pandemic potential and the need to develop vaccines to protect humans from viruses of this subtype. A live attenuated H7N3 virus vaccine was generated by reverse genetics using the HA and NA genes of a low pathogenicity A/chicken/BC/CN-6/04 (H7N3) virus and the six internal protein genes of the cold-adapted A/Ann Arbor/6/60 ca (H2N2) virus. The reassortant H7N3 BC 04 ca vaccine virus was temperature sensitive and showed attenuation in mice and ferrets. Intranasal immunization with one dose of the vaccine protected mice and ferrets when challenged with homologous and heterologous H7 viruses. The reassortant H7N3 BC 04 ca vaccine virus showed comparable levels of attenuation, immunogenicity and efficacy in mice and ferret models. The safety, immunogenicity, and efficacy of this vaccine in mice and ferrets support the evaluation of this vaccine in clinical trials.

  15. CDC Panel Says FluMist Nasal Flu Vaccine Ineffective

    MedlinePlus

    ... 159535.html CDC Panel Says FluMist Nasal Flu Vaccine Ineffective Agency advisors say the product has lost ... without the easier, nasal spray form of flu vaccine next flu season, a panel of experts decided ...

  16. A live attenuated cold adapted influenza A H7N3 virus vaccine provides protection against homologous and heterologous H7 viruses in mice and ferrets

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The appearance of human infections caused by avian influenza A H7 subtype viruses underscore their pandemic potential and the need to develop vaccines to protect humans from viruses of this subtype. A live attenuated H7N3 virus vaccine was generated by reverse genetics using the HA and NA genes of ...

  17. Neandertal cold adaptation: physiological and energetic factors.

    PubMed

    Steegmann, A Theodore; Cerny, Frank J; Holliday, Trenton W

    2002-01-01

    European Neandertals employed a complex set of physiological cold defenses, homologous to those seen in contemporary humans and nonhuman primates. While Neandertal morphological patterns, such as foreshortened extremities and low relative surface-area, may have explained some of the variance in cold resistance, it is suggested the adaptive package was strongly dependent on a rich array of physiological defenses. A summary of the environmental cold conditions in which the Neandertals lived is presented, and a comparative ethnographic model from Tierra del Fuego is used. Muscle and subcutaneous fat are excellent "passive" insulators. Neandertals were quite muscular, but it is unlikely that they could maintain enough superficial body fat to offer much cold protection. A major, high-energy metabolic adaptation facilitated by modest amounts of highly thermogenic brown adipose tissue (BAT) is proposed. In addition, Neandertals would have been protected by general mammalian cold defenses based on systemic vasoconstriction and intensified by acclimatization, aerobic fitness, and localized cold--induced vasodilation. However, these defenses are energetically expensive. Based on contemporary data from circumpolar peoples, it is estimated that Neandertals required 3,360 to 4,480 kcal per day to support strenuous winter foraging and cold resistance costs. Several specific genetic cold adaptations are also proposed--heat shock protein (actually, stress shock protein), an ACP*1 locus somatic growth factor, and a specialized calcium metabolism not as yet understood. PMID:12203812

  18. Seasonal FluMist vaccination induces cross-reactive T cell immunity against H1N1 (2009) influenza and secondary bacterial infections.

    PubMed

    Sun, Keer; Ye, Jianqiang; Perez, Daniel R; Metzger, Dennis W

    2011-01-15

    T cell epitopes have been found to be shared by circulating, seasonal influenza virus strains and the novel pandemic H1N1 influenza infection, but the ability of these common epitopes to provide cross-protection is unknown. We have now directly tested this by examining the ability of live seasonal influenza vaccine (FluMist) to mediate protection against swine-origin H1N1 influenza virus infection. Naive mice demonstrated considerable susceptibility to H1N1 Cal/04/09 infection, whereas FluMist-vaccinated mice had markedly decreased morbidity and mortality. In vivo depletion of CD4(+) or CD8(+) immune cells after vaccination indicated that protective immunity was primarily dependent upon FluMist-induced CD4(+) cells but not CD8(+) T cells. Passive protection studies revealed little role for serum or mucosal Abs in cross-protection. Although H1N1 influenza infection of naive mice induced intensive phagocyte recruitment, pulmonary innate defense against secondary pneumococcal infection was severely suppressed. This increased susceptibility to bacterial infection was correlated with augmented IFN-γ production produced during the recovery stage of H1N1 influenza infection, which was completely suppressed in mice previously immunized with FluMist. Furthermore, susceptibility to secondary bacterial infection was decreased in the absence of type II, but not type I, IFN signaling. Thus, seasonal FluMist treatment not only promoted resistance to pandemic H1N1 influenza infection but also restored innate immunity against complicating secondary bacterial infections.

  19. Current status of live attenuated influenza virus vaccine in the US.

    PubMed

    Belshe, Robert B

    2004-07-01

    The efficacy and effectiveness of cold adapted live attenuated (CAIV-T, FluMist intranasal influenza vaccine is reviewed. CAIV-T consists of approximately 10(7) TCID50 per dose of each influenza A/H1N1, influenza A/H3N2, and influenza B vaccine strain. The exact strains are updated each year to antigenically match the antigens recommended by national health authorities for inclusion in the vaccine. In one year in which the vaccine strain did not well match the epidemic strain, the live attenuated vaccine induced a broad immune response that cross-reacted significantly with the drifted strain. The efficacy of CAIV-T in adults was demonstrated with challenge studies and the effectiveness of the vaccine for reducing febrile upper respiratory illness, days of missed work, and days of antibiotic use was demonstrated in a large field trial. In young children, protective efficacy against culture confirmed influenza was demonstrated in a field trial with overall protective efficacy of 92% during a two year study. Vaccine was also highly protective against a strain not contained in the vaccine, with 86% protective efficacy demonstrated against this significantly drifted virus. Effectiveness measures, including protection against febrile otitis media and visits to the doctor were demonstrated. Live attenuated vaccine provides a significant new tool to help prevent influenza.

  20. Comparison of the safety, vaccine virus shedding, and immunogenicity of influenza virus vaccine, trivalent, types A and B, live cold-adapted, administered to human immunodeficiency virus (HIV)-infected and non-HIV-infected adults.

    PubMed

    King, J C; Treanor, J; Fast, P E; Wolff, M; Yan, L; Iacuzio, D; Readmond, B; O'Brien, D; Mallon, K; Highsmith, W E; Lambert, J S; Belshe, R B

    2000-02-01

    Fifty-seven human immunodeficiency virus (HIV)-infected (CDC class A1-2) and 54 non-HIV-infected adults, not prescreened for influenza susceptibility, were randomized to receive trivalent live attenuated influenza vaccine (LAIV) or placebo intranasally. LAIV was safe and well tolerated with no serious adverse events attributable to vaccine. Reactogenicity rates were similar in LAIV and placebo recipients except that runny nose/nasal congestion was significantly more common in LAIV recipients regardless of HIV status. No prolonged shedding of LAIV was observed in HIV-infected participants. HIV RNA levels were not increased and CD4 counts were not decreased in HIV-infected LAIV recipients compared with placebo recipients after immunization. Shedding of LAIV and increases in antibody titers were infrequent, consistent with prior experience in unscreened adults. The data suggest that inadvertent vaccination with LAIV in relatively asymptomatic HIV-infected adults would not be associated with frequent significant adverse events.

  1. Could human cold adaptation decrease the risk of cardiovascular disease?

    PubMed

    Kralova Lesna, I; Rychlikova, J; Vavrova, L; Vybiral, S

    2015-08-01

    The impact of repeated exposure to cold and cold adaptation on human cardiovascular health is not fully understood. The aim of the study was to determine the effect of cold adaptation on cardiovascular risk factors, thyroid hormones and the capacity of humans to reset the damaging effect of oxidative stress. Ten well cold-adapted winter swimmers (CA) and 16 non-adapted controls (CON) were enroled in this experiment to test whether cold adaptation could influence the parameters of lipoprotein metabolism, cholesterol efflux capacity (CEC), homocysteine, thyroid hormones, antioxidant defence markers (reduced glutathione (GSH), glutathione peroxidase 1 (GPX1), glutathione reductase (GR), catalase (CAT) and paraoxonase 1 (PON1)) and oxidative stress markers (concentration of conjugated dienes (CD)). A decreased apolipoprotein B/apolipoprotein A1 (ApoB/ApoA1) ratio was found in the CA group (p<0.05), but other lipoprotein parameters, including CEC, did not differ significantly. Plasma homocysteine was lower in CA subjects in comparison with controls (p<0.05). Higher triiodothyronine (T3) values were observed in the CA compared to the CON (p<0.05) group, but TSH and other thyroid hormones did not differ between both groups. CA subjects had lower activity of GPX1 (p<0.05), lower concentrations of CD (p<0.05) and increased activities of PON1 (p<0.001) compared to CON subjects. A trend for decreased activity of CAT (p=0.06) in CA compared to CON groups was also observed, but GSH levels did not differ significantly. Zn concentration was higher in the CA group than in the CON group (p<0.001). Human cold adaptation can influence oxidative stress markers. Trends towards the improvement of cardiovascular risk factors in cold-adapted subjects also indicate the positive effect of cold adaptation on cardio-protective mechanisms. PMID:26267514

  2. Protein cold adaptation strategy via a unique seven-amino acid domain in the icefish (Chionodraco hamatus) PEPT1 transporter.

    PubMed

    Rizzello, Antonia; Romano, Alessandro; Kottra, Gabor; Acierno, Raffaele; Storelli, Carlo; Verri, Tiziano; Daniel, Hannelore; Maffia, Michele

    2013-04-23

    Adaptation of organisms to extreme environments requires proteins to work at thermodynamically unfavorable conditions. To adapt to subzero temperatures, proteins increase the flexibility of parts of, or even the whole, 3D structure to compensate for the lower thermal kinetic energy available at low temperatures. This may be achieved through single-site amino acid substitutions in regions of the protein that undergo large movements during the catalytic cycle, such as in enzymes or transporter proteins. Other strategies of cold adaptation involving changes in the primary amino acid sequence have not been documented yet. In Antarctic icefish (Chionodraco hamatus) peptide transporter 1 (PEPT1), the first transporter cloned from a vertebrate living at subzero temperatures, we came upon a unique principle of cold adaptation. A de novo domain composed of one to six repeats of seven amino acids (VDMSRKS), placed as an extra stretch in the cytosolic COOH-terminal region, contributed per se to cold adaptation. VDMSRKS was in a protein region uninvolved in transport activity and, notably, when transferred to the COOH terminus of a warm-adapted (rabbit) PEPT1, it conferred cold adaptation to the receiving protein. Overall, we provide a paradigm for protein cold adaptation that relies on insertion of a unique domain that confers greater affinity and maximal transport rates at low temperatures. Due to its ability to transfer a thermal trait, the VDMSRKS domain represents a useful tool for future cell biology or biotechnological applications. PMID:23569229

  3. Anti-Biofilm Activities from Marine Cold Adapted Bacteria Against Staphylococci and Pseudomonas aeruginosa

    PubMed Central

    Papa, Rosanna; Selan, Laura; Parrilli, Ermenegilda; Tilotta, Marco; Sannino, Filomena; Feller, Georges; Tutino, Maria L.; Artini, Marco

    2015-01-01

    Microbial biofilms have great negative impacts on the world’s economy and pose serious problems to industry, public health and medicine. The interest in the development of new approaches for the prevention and treatment of bacterial adhesion and biofilm formation has increased. Since, bacterial pathogens living in biofilm induce persistent chronic infections due to the resistance to antibiotics and host immune system. A viable approach should target adhesive properties without affecting bacterial vitality in order to avoid the appearance of resistant mutants. Many bacteria secrete anti-biofilm molecules that function in regulating biofilm architecture or mediating the release of cells from it during the dispersal stage of biofilm life cycle. Cold-adapted marine bacteria represent an untapped reservoir of biodiversity able to synthesize a broad range of bioactive compounds, including anti-biofilm molecules. The anti-biofilm activity of cell-free supernatants derived from sessile and planktonic cultures of cold-adapted bacteria belonging to Pseudoalteromonas, Psychrobacter, and Psychromonas species were tested against Staphylococcus aureus, Staphylococcus epidermidis, and Pseudomonas aeruginosa strains. Reported results demonstrate that we have selected supernatants, from cold-adapted marine bacteria, containing non-biocidal agents able to destabilize biofilm matrix of all tested pathogens without killing cells. A preliminary physico-chemical characterization of supernatants was also performed, and these analyses highlighted the presence of molecules of different nature that act by inhibiting biofilm formation. Some of them are also able to impair the initial attachment of the bacterial cells to the surface, thus likely containing molecules acting as anti-biofilm surfactant molecules. The described ability of cold-adapted bacteria to produce effective anti-biofilm molecules paves the way to further characterization of the most promising molecules and to test their

  4. Anti-Biofilm Activities from Marine Cold Adapted Bacteria Against Staphylococci and Pseudomonas aeruginosa.

    PubMed

    Papa, Rosanna; Selan, Laura; Parrilli, Ermenegilda; Tilotta, Marco; Sannino, Filomena; Feller, Georges; Tutino, Maria L; Artini, Marco

    2015-01-01

    Microbial biofilms have great negative impacts on the world's economy and pose serious problems to industry, public health and medicine. The interest in the development of new approaches for the prevention and treatment of bacterial adhesion and biofilm formation has increased. Since, bacterial pathogens living in biofilm induce persistent chronic infections due to the resistance to antibiotics and host immune system. A viable approach should target adhesive properties without affecting bacterial vitality in order to avoid the appearance of resistant mutants. Many bacteria secrete anti-biofilm molecules that function in regulating biofilm architecture or mediating the release of cells from it during the dispersal stage of biofilm life cycle. Cold-adapted marine bacteria represent an untapped reservoir of biodiversity able to synthesize a broad range of bioactive compounds, including anti-biofilm molecules. The anti-biofilm activity of cell-free supernatants derived from sessile and planktonic cultures of cold-adapted bacteria belonging to Pseudoalteromonas, Psychrobacter, and Psychromonas species were tested against Staphylococcus aureus, Staphylococcus epidermidis, and Pseudomonas aeruginosa strains. Reported results demonstrate that we have selected supernatants, from cold-adapted marine bacteria, containing non-biocidal agents able to destabilize biofilm matrix of all tested pathogens without killing cells. A preliminary physico-chemical characterization of supernatants was also performed, and these analyses highlighted the presence of molecules of different nature that act by inhibiting biofilm formation. Some of them are also able to impair the initial attachment of the bacterial cells to the surface, thus likely containing molecules acting as anti-biofilm surfactant molecules. The described ability of cold-adapted bacteria to produce effective anti-biofilm molecules paves the way to further characterization of the most promising molecules and to test their

  5. Cold-adapted proteases as an emerging class of therapeutics.

    PubMed

    Fornbacke, Marcus; Clarsund, Mats

    2013-06-01

    Proteases have been used in medicine for several decades and are an established and well tolerated class of therapeutic agent. These proteases were sourced from mammals or bacteria that exist or have adapted to moderate temperatures (mesophilic organisms); however, proteases derived from organisms from cold environments-cold-adapted or psychrophilic proteases-generally have high specific activity, low substrate affinity, and high catalytic rates at low and moderate temperatures. Made possible by greater flexibility, psychrophilic enzymes interact with and transform the substrate at lower energy costs. Cold-adapted proteases have been used in a wide range of applications, including industrial functions, textiles, cleaning/hygiene products, molecular biology, environmental bioremediations, consumer food products, cosmetics, and pharmaceutical production. In addition to these applications, they have also shown promise as therapeutic modalities for cosmeceutical applications (by reducing glabellar [frown] lines) and a number of disease conditions, including bacterial infections (by disrupting biofilms to prevent bacterial infection), topical wound management (when used as a debridement agent to remove necrotic tissue and fibrin clots), oral/dental health management (by removing plaque and preventing periodontal disease), and in viral infections (by reducing the infectivity of viruses, such as human rhinovirus 16 and herpes simplex virus). Psychrophilic proteases with greater activity and stability (than the original organism-derived variant) have been developed; this coupled with available manufacturing recombinant production techniques suggests that cold-adapted proteases have a promising future as a distinct therapeutic class with diverse clinical applications.

  6. Cold-adapted proteases as an emerging class of therapeutics.

    PubMed

    Fornbacke, Marcus; Clarsund, Mats

    2013-06-01

    Proteases have been used in medicine for several decades and are an established and well tolerated class of therapeutic agent. These proteases were sourced from mammals or bacteria that exist or have adapted to moderate temperatures (mesophilic organisms); however, proteases derived from organisms from cold environments-cold-adapted or psychrophilic proteases-generally have high specific activity, low substrate affinity, and high catalytic rates at low and moderate temperatures. Made possible by greater flexibility, psychrophilic enzymes interact with and transform the substrate at lower energy costs. Cold-adapted proteases have been used in a wide range of applications, including industrial functions, textiles, cleaning/hygiene products, molecular biology, environmental bioremediations, consumer food products, cosmetics, and pharmaceutical production. In addition to these applications, they have also shown promise as therapeutic modalities for cosmeceutical applications (by reducing glabellar [frown] lines) and a number of disease conditions, including bacterial infections (by disrupting biofilms to prevent bacterial infection), topical wound management (when used as a debridement agent to remove necrotic tissue and fibrin clots), oral/dental health management (by removing plaque and preventing periodontal disease), and in viral infections (by reducing the infectivity of viruses, such as human rhinovirus 16 and herpes simplex virus). Psychrophilic proteases with greater activity and stability (than the original organism-derived variant) have been developed; this coupled with available manufacturing recombinant production techniques suggests that cold-adapted proteases have a promising future as a distinct therapeutic class with diverse clinical applications. PMID:25135820

  7. The influence of the social thermoregulation on the cold-adaptive growth of BAT in hairless and furred mice.

    PubMed

    Heldmaier, G

    1975-03-26

    When mice were living in groups they developed less brown adipose tissue (BAT) during cold adaptation as compared with single mice. This effect of social aggregation was more pronounced in genetically hairless mice than in furred mice. In both races of mice the most significant difference in BAT growth was found between single mice and pairs of mice, indicating that the formation of pairs causes the relatively most effective improvement of thermal balance.

  8. Highly phosphorylated core oligosaccharide structures from cold-adapted Psychromonas arctica.

    PubMed

    Corsaro, Maria M; Pieretti, Giuseppina; Lindner, Buko; Lanzetta, Rosa; Parrilli, Ermenegilda; Tutino, Maria L; Parrilli, Michelangelo

    2008-01-01

    Many cold habitats contain plenty of microorganisms that represent the most abundant cold-adapted life forms on earth. These organisms have developed a wide range of adaptations that involve the cell wall of the microorganism. In particular, bacteria enhance the synthesis of unsaturated fatty acids of membrane lipids to maintain the membrane fluidity, but very little is known about the adaptational changes in the structure of the lipopolysaccharides (LPSs), the main constituent of the outer leaflet of the outer membrane of Gram-negative bacteria. The aim of this study was to investigate the chemical structure of these LPSs for insight into the temperature-adaptation mechanism. For this objective, the cold-adapted Psychromonas arctica bacterium, which lives in the arctic sea-water near Spitzbergen (Svalbard islands, Arctic) was cultivated at 4 degrees C. The lipooligosaccharides (LOSs) were isolated and analysed by means of chemical analysis and electrospray ionisation high-resolution Fourier transform mass spectrometry. The LOS was then degraded either by mild hydrazinolysis (O-deacylation) or with hot 4 M KOH (N-deacylation). Both products were investigated in detail by using 1H and 13C NMR spectroscopy and mass spectrometry. The core consists of a mixture of species that differ because of the presence of nonstoichiometric D-fructose and/or D-galacturonic acid units.

  9. Cutting edge: rapid boosting of cross-reactive memory CD8 T cells broadens the protective capacity of the Flumist vaccine.

    PubMed

    Slütter, Bram; Pewe, Lecia L; Lauer, Peter; Harty, John T

    2013-04-15

    Memory CD8 T cells recognizing conserved proteins from influenza A virus (IAV), such as nucleoprotein, have the potential to provide protection in individuals who lack the proper neutralizing Abs. In this study, we show that the most potent CD8 T cell-inducing influenza vaccine on the market (Flumist) does not induce sufficient numbers of cross-reactive CD8 T cells to provide substantial protection against lethal nonhomologous IAV challenge. However, Flumist-primed CD8 T cells rapidly acquire memory characteristics and can respond to short-interval boosting to greatly enlarge the IAV-specific memory pool, which is sufficient to protect mice from nonhomologous IAV challenge. Thus, a current vaccine strategy, Flumist, may serve as a priming platform for the rapid induction of large numbers of memory CD8 T cells with the capacity for broad protection against influenza.

  10. Rapid boosting of cross-reactive memory CD8 T-cells broadens the protective capacity of the Flumist® vaccine

    PubMed Central

    Slütter, Bram; Pewe, Lecia L.; Lauer, Peter; Harty, John T.

    2013-01-01

    Memory CD8 T-cells recognizing conserved proteins from influenza A virus (IAV) such as nucleoprotein (NP), have the potential to provide protection in individuals who lack the proper neutralizing antibodies. Here we show that the most potent CD8 T-cell inducing influenza vaccine on the market (Flumist®), does not induce sufficient numbers of cross-reactive CD8 T-cells to provide substantial protection against lethal non-homologous IAV challenge. However, Flumist primed CD8 T-cells rapidly acquire memory characteristics and can respond to short interval boosting to greatly enlarge the IAV specific memory pool, sufficient to protect mice from non-homologous IAV challenge. Thus, a current vaccine strategy, Flumist, may serve as a priming platform for rapid induction of large numbers of memory CD8 T-cells with the capacity for broad protection against influenza. PMID:23467935

  11. Generation and protective efficacy of a cold-adapted attenuated avian H9N2 influenza vaccine.

    PubMed

    Wei, Yandi; Qi, Lu; Gao, Huijie; Sun, Honglei; Pu, Juan; Sun, Yipeng; Liu, Jinhua

    2016-01-01

    To prevent H9N2 avian influenza virus infection in chickens, a long-term vaccination program using inactivated vaccines has been implemented in China. However, the protective efficacy of inactivated vaccines against antigenic drift variants is limited, and H9N2 influenza virus continues to circulate in vaccinated chicken flocks in China. Therefore, developing a cross-reactive vaccine to control the impact of H9N2 influenza in the poultry industry remains a high priority. In the present study, we developed a live cold-adapted H9N2 influenza vaccine candidate (SD/01/10-ca) by serial passages in embryonated eggs at successively lower temperatures. A total of 13 amino acid mutations occurred during the cold-adaptation of this H9N2 virus. The candidate was safe in chickens and induced robust hemagglutination-inhibition antibody responses and influenza virus-specific CD4(+) and CD8(+) T cell immune responses in chickens immunized intranasally. Importantly, the candidate could confer protection of chickens from homologous and heterogenous H9N2 viruses. These results demonstrated that the cold-adapted attenuated H9N2 virus would be selected as a vaccine to control the infection of prevalent H9N2 influenza viruses in chickens. PMID:27457755

  12. Generation and protective efficacy of a cold-adapted attenuated avian H9N2 influenza vaccine

    PubMed Central

    Wei, Yandi; Qi, Lu; Gao, Huijie; Sun, Honglei; Pu, Juan; Sun, Yipeng; Liu, Jinhua

    2016-01-01

    To prevent H9N2 avian influenza virus infection in chickens, a long-term vaccination program using inactivated vaccines has been implemented in China. However, the protective efficacy of inactivated vaccines against antigenic drift variants is limited, and H9N2 influenza virus continues to circulate in vaccinated chicken flocks in China. Therefore, developing a cross-reactive vaccine to control the impact of H9N2 influenza in the poultry industry remains a high priority. In the present study, we developed a live cold-adapted H9N2 influenza vaccine candidate (SD/01/10-ca) by serial passages in embryonated eggs at successively lower temperatures. A total of 13 amino acid mutations occurred during the cold-adaptation of this H9N2 virus. The candidate was safe in chickens and induced robust hemagglutination-inhibition antibody responses and influenza virus–specific CD4+ and CD8+ T cell immune responses in chickens immunized intranasally. Importantly, the candidate could confer protection of chickens from homologous and heterogenous H9N2 viruses. These results demonstrated that the cold-adapted attenuated H9N2 virus would be selected as a vaccine to control the infection of prevalent H9N2 influenza viruses in chickens. PMID:27457755

  13. Genomic and Expression Analyses of Cold-Adapted Microorganisms.

    SciTech Connect

    Bakermans, Corien; Bergholz, Peter W.; Rodrigues, Debora F.; Vishnivetskaya, T.; Ayala-del-Río, Hector L.; Tiedje, James M.

    2011-01-01

    Contents 7.1 Introduction 7.2 Ecological evidence of bacterial adaptation to cold 7.2.1 Characteristics of cold environments and implications for microbial ecology 7.2.2 Ecological adaptation in Exiguobacterium spp. and Psychrobacter spp. 7.3 Gene Expression Responses to the Cold 7.3.1 Fundamentals of Gene Expression Responses to Cold 7.3.2 Acclimation for Life in Cold Habitats 7.3.2.1 Translation and Chaperone Proteins: Safeguarding the functional units of cellular physiology 7.3.2.2 Carbon and Energy metabolism: resource efficiency over long generation times 7.3.2.3 Amino Acid Biosynthesis: Species-specific responses to species-specific deficiencies 7.3.2.4 Compatible solutes: a concomitant response in cryoenvironments 7.3.2.5 Membrane fluidity: A major role in the overall metabolic rate at temperature 7.3.2.6 The cell wall at low temperature: A poorly understood growth rate determinant 7.3.2.7 Transporters: The balance between local nutrient uptake and depletion 7.3.2.8 Genome plasticity. The potential role of transposases and repeated sequences. 7.4 Protein adaptations to cold 7.4.1 The low temperature challenge 7.4.2 The stability activity relationship 7.4.3 Structural features of cold adapted enzymes. 7.4.4 Hydrophobic interactions 7.4.5 Electrostatic interactions 7.4.5.1 Arginine 7.4.5.2 Acidic residues 7.4.6 Structural elements 7.4.6.1 -helices and -sheets 7.4.6.2 Proline and Glycine 7.4.6.3 Disordered regions 7.5 Comparison of cold- and warm-adapted Exiguobacterium strains 7.5.1 Phylogeny reflects adaptations to environmental conditions 7.5.2 Genomic comparison of two strains 7.6 Summary and future directions

  14. [Bioremediation of petroleum hydrocarbon-contaminated soils by cold-adapted microorganisms: research advance].

    PubMed

    Wang, Shi-jie; Wang, Xiang; Lu, Gui-lan; Wang, Qun-hui; Li, Fa-sheng; Guo, Guan-lin

    2011-04-01

    Cold-adapted microorganisms such as psychrotrophs and psychrophiles widely exist in the soils of sub-Arctic, Arctic, Antarctic, alpine, and high mountains, being the important microbial resources for the biodegradation of petroleum hydrocarbons at low temperature. Using the unique advantage of cold-adapted microorganisms to the bioremediation of petroleum hydrocarbon-contaminated soils in low temperature region has become a research hotspot. This paper summarized the category and cold-adaptation mechanisms of the microorganisms able to degrade petroleum hydrocarbon at low temperature, biodegradation characteristics and mechanisms of different petroleum fractions under the action of cold-adapted microorganisms, bio-stimulation techniques for improving biodegradation efficiency, e. g., inoculating petroleum-degrading microorganisms and adding nutrients or bio-surfactants, and the present status of applying molecular biotechnology in this research field, aimed to provide references to the development of bioremediation techniques for petroleum hydrocarbon-contaminated soils.

  15. Searching for signatures of cold adaptations in modern and archaic humans: hints from the brown adipose tissue genes.

    PubMed

    Sazzini, M; Schiavo, G; De Fanti, S; Martelli, P L; Casadio, R; Luiselli, D

    2014-09-01

    Adaptation to low temperatures has been reasonably developed in the human species during the colonization of the Eurasian landmass subsequent to Out of Africa migrations of anatomically modern humans. In addition to morphological and cultural changes, also metabolic ones are supposed to have favored human isolation from cold and body heat production and this can be hypothesized also for most Neandertal and at least for some Denisovan populations, which lived in geographical areas that strongly experienced the last glacial period. Modulation of non-shivering thermogenesis, for which adipocytes belonging to the brown adipose tissue are the most specialized cells, might have driven these metabolic adaptations. To perform an exploratory analysis aimed at looking into this hypothesis, variation at 28 genes involved in such functional pathway was investigated in modern populations from different climate zones, as well as in Neandertal and Denisovan genomes. Patterns of variation at the LEPR gene, strongly related to increased heat dissipation by mitochondria, appeared to have been shaped by positive selection in modern East Asians, but not in Europeans. Moreover, a single potentially cold-adapted LEPR allele, different from the supposed adaptive one identified in Homo sapiens, was found also in Neandertal and Denisovan genomes. These findings suggest that independent mechanisms for cold adaptations might have been developed in different non-African human groups, as well as that the evolution of possible enhanced thermal efficiency in Neandertals and in some Denisovan populations has plausibly entailed significant changes also in other functional pathways than in the examined one.

  16. Searching for signatures of cold adaptations in modern and archaic humans: hints from the brown adipose tissue genes

    PubMed Central

    Sazzini, M; Schiavo, G; De Fanti, S; Martelli, P L; Casadio, R; Luiselli, D

    2014-01-01

    Adaptation to low temperatures has been reasonably developed in the human species during the colonization of the Eurasian landmass subsequent to Out of Africa migrations of anatomically modern humans. In addition to morphological and cultural changes, also metabolic ones are supposed to have favored human isolation from cold and body heat production and this can be hypothesized also for most Neandertal and at least for some Denisovan populations, which lived in geographical areas that strongly experienced the last glacial period. Modulation of non-shivering thermogenesis, for which adipocytes belonging to the brown adipose tissue are the most specialized cells, might have driven these metabolic adaptations. To perform an exploratory analysis aimed at looking into this hypothesis, variation at 28 genes involved in such functional pathway was investigated in modern populations from different climate zones, as well as in Neandertal and Denisovan genomes. Patterns of variation at the LEPR gene, strongly related to increased heat dissipation by mitochondria, appeared to have been shaped by positive selection in modern East Asians, but not in Europeans. Moreover, a single potentially cold-adapted LEPR allele, different from the supposed adaptive one identified in Homo sapiens, was found also in Neandertal and Denisovan genomes. These findings suggest that independent mechanisms for cold adaptations might have been developed in different non-African human groups, as well as that the evolution of possible enhanced thermal efficiency in Neandertals and in some Denisovan populations has plausibly entailed significant changes also in other functional pathways than in the examined one. PMID:24667833

  17. In vivo assessment of NS1-truncated influenza virus with a novel SLSYSINWRH motif as a self-adjuvanting live attenuated vaccine.

    PubMed

    Ngunjiri, John M; Ali, Ahmed; Boyaka, Prosper; Marcus, Philip I; Lee, Chang-Won

    2015-01-01

    Mutants of influenza virus that encode C-terminally truncated NS1 proteins (NS1-truncated mutants) characteristically induce high interferon responses. The dual activity of interferon in blocking virus replication and enhancing the development of adaptive immune responses makes these mutants promising as self-adjuvanting live-attenuated influenza vaccine (LAIV) candidates. Yet, among the NS1-truncated mutants, the length of NS1 is not directly correlated with the interferon-inducing efficiency, the level of attenuation, or effectiveness as LAIV. Using quantitative in vitro biologically active particle subpopulation analysis as a tool to identify potential LAIV candidates from a pool of NS1-truncated mutants, we previously predicted that a NS1-truncated mutant pc2, which was less effective as a LAIV in chickens, would be sufficiently effective as a LAIV in mammalian hosts. In this study, we confirmed that pc2 protected mice and pigs against heterologous virus challenge in terms of preventing clinical signs and reducing virus shedding. pc2 expresses a unique SLSYSINWRH motif at the C-terminus of its truncated NS1. Deletion of the SLSYSINWRH motif led to ~821-fold reduction in the peak yield of type I interferon induced in murine cells. Furthermore, replacement of the SLSYSINWRH motif with the wildtype MVKMDQAIMD sequence did not restore the interferon-inducing efficiency. The diminished interferon induction capacity in the absence of the SLSYSINWRH motif was similar to that observed in other mutants which are less effective LAIV candidates. Remarkably, pc2 induced 16-fold or more interferon in human lung and monkey kidney cells compared to the temperature-sensitive, cold-adapted Ann Arbor virus that is currently used as a master backbone for LAIVs such as FluMist. Although the mechanism by which the SLSYSINWRH motif regulates the vaccine properties of pc2 has not been elucidated, this motif has potential use in engineering self-adjuvanting NS1-truncated-based LAIVs.

  18. Mechanism for stabilizing mRNAs involved in methanol-dependent methanogenesis of cold-adaptive Methanosarcina mazei zm-15.

    PubMed

    Cao, Yi; Li, Jie; Jiang, Na; Dong, Xiuzhu

    2014-02-01

    Methylotrophic methanogenesis predominates at low temperatures in the cold Zoige wetland in Tibet. To elucidate the basis of cold-adapted methanogenesis in these habitats, Methanosarcina mazei zm-15 was isolated, and the molecular basis of its cold activity was studied. For this strain, aceticlastic methanogenesis was reduced 7.7-fold during growth at 15°C versus 30°C. Methanol-derived methanogenesis decreased only 3-fold under the same conditions, suggesting that it is more cold adaptive. Reverse transcription-quantitative PCR (RT-qPCR) detected <2-fold difference in the transcript abundances of mtaA1, mtaB1, and mtaC1, the methanol methyltransferase (Mta) genes, in 30°C versus 15°C culture, while ackA and pta mRNAs, encoding acetate kinase (Ack) and phosphotransacetylase (Pta) in aceticlastic methanogenesis, were 4.5- and 6.8-fold higher in 30°C culture than in 15°C culture. The in vivo half-lives of mtaA1 and mtaC1B1 mRNAs were similar in 30°C and 15°C cultures. However, the pta-ackA mRNA half-life was significantly reduced in 15°C culture compared to 30°C culture. Using circularized RNA RT-PCR, large 5' untranslated regions (UTRs) (270 nucleotides [nt] and 238 nt) were identified for mtaA1 and mtaC1B1 mRNAs, while only a 27-nt 5' UTR was present in the pta-ackA transcript. Removal of the 5' UTRs significantly reduced the in vitro half-lives of mtaA1 and mtaC1B1 mRNAs. Remarkably, fusion of the mtaA1 or mtaC1B1 5' UTRs to pta-ackA mRNA increased its in vitro half-life at both 30°C and 15°C. These results demonstrate that the large 5' UTRs significantly enhance the stability of the mRNAs involved in methanol-derived methanogenesis in the cold-adaptive M. mazei zm-15.

  19. Behavioral buffering of global warming in a cold-adapted lizard.

    PubMed

    Ortega, Zaida; Mencía, Abraham; Pérez-Mellado, Valentín

    2016-07-01

    Alpine lizards living in restricted areas might be particularly sensitive to climate change. We studied thermal biology of Iberolacerta cyreni in high mountains of central Spain. Our results suggest that I. cyreni is a cold-adapted thermal specialist and an effective thermoregulator. Among ectotherms, thermal specialists are more threatened by global warming than generalists. Alpine lizards have no chance to disperse to new suitable habitats. In addition, physiological plasticity is unlikely to keep pace with the expected rates of environmental warming. Thus, lizards might rely on their behavior in order to deal with ongoing climate warming. Plasticity of thermoregulatory behavior has been proposed to buffer the rise of environmental temperatures. Therefore, we studied the change in body and environmental temperatures, as well as their relationships, for I. cyreni between the 1980s and 2012. Air temperatures have increased more than 3.5°C and substrate temperatures have increased by 6°C in the habitat of I. cyreni over the last 25 years. However, body temperatures of lizards have increased less than 2°C in the same period, and the linear relationship between body and environmental temperatures remains similar. These results show that alpine lizards are buffering the potential impact of the increase in their environmental temperatures, most probably by means of their behavior. Body temperatures of I. cyreni are still cold enough to avoid any drop in fitness. Nonetheless, if warming continues, behavioral buffering might eventually become useless, as it would imply spending too much time in shelter, losing feeding, and mating opportunities. Eventually, if body temperature exceeds the thermal optimum in the near future, fitness would decrease abruptly.

  20. Behavioral buffering of global warming in a cold-adapted lizard.

    PubMed

    Ortega, Zaida; Mencía, Abraham; Pérez-Mellado, Valentín

    2016-07-01

    Alpine lizards living in restricted areas might be particularly sensitive to climate change. We studied thermal biology of Iberolacerta cyreni in high mountains of central Spain. Our results suggest that I. cyreni is a cold-adapted thermal specialist and an effective thermoregulator. Among ectotherms, thermal specialists are more threatened by global warming than generalists. Alpine lizards have no chance to disperse to new suitable habitats. In addition, physiological plasticity is unlikely to keep pace with the expected rates of environmental warming. Thus, lizards might rely on their behavior in order to deal with ongoing climate warming. Plasticity of thermoregulatory behavior has been proposed to buffer the rise of environmental temperatures. Therefore, we studied the change in body and environmental temperatures, as well as their relationships, for I. cyreni between the 1980s and 2012. Air temperatures have increased more than 3.5°C and substrate temperatures have increased by 6°C in the habitat of I. cyreni over the last 25 years. However, body temperatures of lizards have increased less than 2°C in the same period, and the linear relationship between body and environmental temperatures remains similar. These results show that alpine lizards are buffering the potential impact of the increase in their environmental temperatures, most probably by means of their behavior. Body temperatures of I. cyreni are still cold enough to avoid any drop in fitness. Nonetheless, if warming continues, behavioral buffering might eventually become useless, as it would imply spending too much time in shelter, losing feeding, and mating opportunities. Eventually, if body temperature exceeds the thermal optimum in the near future, fitness would decrease abruptly. PMID:27386098

  1. [Cloning and expression of endoglucanase of marine cold-adapted bacteria Pseudoalteromonas sp. MB-1].

    PubMed

    You, Yin-wei; Wang, Tian-hong

    2005-02-01

    The cold-adapted gram-negative rod bacterium MB-1 which could secret cellulase was screened from mud of the bottom of the Huanghai. According to the sequence of 16S rDNA, this bacterium screened was identified as one species of Pseudoalteromonas and was named as Pseudoalteromonas sp. MB-1. The gene celA encoding cold-adapted endogluanase was cloned and then jointed to pGEX-4T-1 to construct expression plasmid pGEX-celA which was expressed in E. coli BL21. Analysis to the supernatant of E. coli sonicate revealed that the concentration of GST-CelA was about 78.5 mg/L. Properties of the fusion enzyme of GST-CelA including the optimum temperature at 35 degrees C and the optimum pH about 7.2, showed that this fusion enzyme still belonged to cold-adapted enzyme and neutral enzyme. The result lays solid base for the fundamental theory and application research on cold-adapted cellulase from Pseudoalteromonas sp. MB-1.

  2. The Lysozyme from Insect (Manduca sexta) is a Cold-Adapted Enzyme

    SciTech Connect

    Sotelo-Mundo,R.; Lopez-Zavala, A.; Garcia-Orozco, K.; Arvizu-Flores, A.; Velazquez-Contreras, E.; Valenzuela-Soto, E.; Rojo-Dominguez, A.; Kanost, M.

    2007-01-01

    Enzymatic activity is dependent on temperature, although some proteins have evolved to retain activity at low temperatures at the expense of stability. Cold adapted enzymes are present in a variety of organisms and there is ample interest in their structure-function relationships. Lysozyme (E.C. 3.2.1.17) is one of the most studied enzymes due to its antibacterial activity against Gram positive bacteria and is also a cold adapted protein. In this work the characterization of lysozyme from the insect Manduca sexta and its activity at low temperatures is presented. Both M. sexta lysozymes natural and recombinant showed a higher content of {alpha}-helix secondary structure compared to that of hen egg white lysozyme and a higher specific enzymatic activity in the range of 5-30 {sup o}C. These results together with measured thermodynamic activation parameters support the designation of M. sexta lysozyme as a cold adapted enzyme. Therefore, the insect recombinant lysozyme is feasible as a model for structure-function studies for cold-adapted proteins.

  3. The lysozyme from insect (Manduca sexta) is a cold-adapted enzyme

    PubMed Central

    Sotelo-Mundo, Rogerio R.; Lopez-Zavala, Alonso A.; Garcia-Orozco, Karina D.; Arvizu-Flores, Aldo A.; Velazquez-Contreras, Enrique F.; Valenzuela-Soto, Elisa M.; Rojo-Dominguez, Arturo; Kanost, Michael R.

    2008-01-01

    Enzymatic activity is dependent on temperature, although some proteins have evolved to retain activity at low temperatures at the expense of stability. Cold adapted enzymes are present in a variety of organisms and there is ample interest in their structure-function relationships. Lysozyme (E.C. 3.2.1.17) is one of the most studied enzymes due to its antibacterial activity against Gram positive bacteria and is also a cold adapted protein. In this work the characterization of lysozyme from the insect Manduca sexta and its activity at low temperatures is presented. Both M. sexta lysozymes natural and recombinant showed a higher content of α-helix secondary structure compared to that of hen egg white lysozyme and a higher specific enzymatic activity in the range of 5−30 °C. These results together with measured thermodynamical activation parameters support the designation of M. sexta lysozyme as a cold adapted enzyme. Therefore, the insect recombinant lysozyme is feasible as a model for structure-function studies for cold-adapted proteins. PMID:17979817

  4. Protein cold adaptation: Role of physico-chemical parameters in adaptation of proteins to low temperatures.

    PubMed

    Shokrollahzade, Soheila; Sharifi, Fatemeh; Vaseghi, Akbar; Faridounnia, Maryam; Jahandideh, Samad

    2015-10-21

    During years 2007 and 2008, we published three papers (Jahandideh, 2007a, JTB, 246, 159-166; Jahandideh, 2007b, JTB, 248, 721-726; Jahandideh, 2008, JTB, 255, 113-118) investigating sequence and structural parameters in adaptation of proteins to low temperatures. Our studies revealed important features in cold-adaptation of proteins. Here, we calculate values of a new set of physico-chemical parameters and perform a comparative systematic analysis on a more comprehensive database of psychrophilic-mesophilic homologous protein pairs. Our obtained results confirm that psychrophilicity rules are not merely the inverse rules of thermostability; for instance, although contact order is reported as a key feature in thermostability, our results have shown no significant difference between contact orders of psychrophilic proteins compared to mesophilic proteins. We are optimistic that these findings would help future efforts to propose a strategy for designing cold-adapted proteins.

  5. Recombinant cold-adapted trypsin I from Atlantic cod-expression, purification, and identification.

    PubMed

    Jónsdóttir, Gudrún; Bjarnason, Jón Bragi; Gudmundsdóttir, Agústa

    2004-01-01

    Atlantic cod trypsin I is a cold-adapted proteolytic enzyme exhibiting approximately 20 times higher catalytic efficiency (kcat/KM) than its mesophilic bovine counterpart for the simple amide substrate BAPNA. In general, cold-adapted proteolytic enzymes are sensitive to autolytic degradation, thermal inactivation as well as molecular aggregation, even at temperatures as low as 18-25 degrees C which may explain the problems observed with their expression, activation, and purification. Prior to the data presented here, there have been no reports in the literature on the expression of psychrophilic or cold-adapted proteolytic enzymes from fish. Nevertheless, numerous cold-adapted proteolytic microbial enzymes have been successfully expressed in bacteria and yeast. This report describes successful expression, activation, and purification of the recombinant cod trypsin I in the His-Patch ThioFusion Escherichia coli expression system. The E. coli pThioHis expression vector used in the study enabled the formation of a fusion protein between a highly soluble fraction of HP-thioredoxin contained in the vector and the N-terminal end of the precursor form of cod trypsin I. The HP-thioredoxin part of the fusion protein binds to a metal-chelating ProBond column, which facilitated its purification. The cod trypsin I part of the purified fusion protein was released by proteolytic cleavage, resulting in concomitant activation of the recombinant enzyme. The recombinant cod trypsin I was purified to homogeneity on a trypsin-specific benzamidine affinity column. The identity of the recombinant enzyme was demonstrated by electrophoresis and chromatography.

  6. [Gene cloning, expression and characterization of two cold-adapted lipases from Penicillium sp. XMZ-9].

    PubMed

    Zheng, Xiaomei; Wu, Ningfeng; Fan, Yunliu

    2012-04-01

    Cold-adapted lipases are attractive biocatalysts that can be used at low temperatures as additives in food products, laundry detergents, and the organic synthesis of chiral intermediates. Cold-adapted lipases are normally found in microorganisms that survive at low temperatures. A fungi strain XMZ-9 exhibiting lipolytic activity was isolated from the soil of glaciers in Xinjiang by the screening plates using 1% tributyrin as the substrate and Victoria blue as an indicator. Based on morphological characteristics and phylogenetic comparisons of its 18S rDNA, the strain was identified as Penicillium sp. The partial nucleotide sequences of these two lipase related genes, LipA and LipB, were obtained by touchdown PCR using the degenerate primers designed according to the conservative domains of lipase. The full-length sequences of two genes were obtained by genome walking. The gene lipA contained 1 014 nucleotides, without any intron, comprising one open reading frame encoding a polypeptide of 337 amino acids. The gene lipB comprised two introns (61 bp and 49 bp) and a coding region sequence of 1 122 bp encoding a polypeptide of 373 amino acids, cDNA sequences of both lipA and lipB were cloned and expressed in Escherichia coli BL21 (DE3). The recombinant LipA was mostly expressed as inclusion bodies, and recovered lipase activity at low temperature after in vitro refolded by dilution. Differently, the recombinant LipB was expressed in the soluble form and then purified by Ni-NTA affinity chromatography Column. It showed high lipase activity at low temperature. These results indicated that they were cold-adapted enzymes. This study paves the way for the further research of these cold-adapted lipases for application in the industry.

  7. Cold Adaptation, Ca2+ Dependency and Autolytic Stability Are Related Features in a Highly Active Cold-Adapted Trypsin Resistant to Autoproteolysis Engineered for Biotechnological Applications

    PubMed Central

    Olivera-Nappa, Alvaro; Reyes, Fernando; Andrews, Barbara A.; Asenjo, Juan A.

    2013-01-01

    Pig trypsin is routinely used as a biotechnological tool, due to its high specificity and ability to be stored as an inactive stable zymogen. However, it is not an optimum enzyme for conditions found in wound debriding for medical uses and trypsinization processes for protein analysis and animal cell culturing, where low Ca2+ dependency, high activity in mild conditions and easy inactivation are crucial. We isolated and thermodynamically characterized a highly active cold-adapted trypsin for medical and laboratory use that is four times more active than pig trypsin at 10° C and at least 50% more active than pig trypsin up to 50° C. Contrary to pig trypsin, this enzyme has a broad optimum pH between 7 and 10 and is very insensitive to Ca2+ concentration. The enzyme is only distantly related to previously described cryophilic trypsins. We built and studied molecular structure models of this trypsin and performed molecular dynamic calculations. Key residues and structures associated with calcium dependency and cryophilicity were identified. Experiments indicated that the protein is unstable and susceptible to autoproteolysis. Correlating experimental results and structural predictions, we designed mutations to improve the resistance to autoproteolysis and conserve activity for longer periods after activation. One single mutation provided around 25 times more proteolytic stability. Due to its cryophilic nature, this trypsin is easily inactivated by mild denaturation conditions, which is ideal for controlled proteolysis processes without requiring inhibitors or dilution. We clearly show that cold adaptation, Ca2+ dependency and autolytic stability in trypsins are related phenomena that are linked to shared structural features and evolve in a concerted fashion. Hence, both structurally and evolutionarily they cannot be interpreted and studied separately as previously done. PMID:23951314

  8. Cold adaptation increases rates of nutrient flow and metabolic plasticity during cold exposure in Drosophila melanogaster.

    PubMed

    Williams, Caroline M; McCue, Marshall D; Sunny, Nishanth E; Szejner-Sigal, Andre; Morgan, Theodore J; Allison, David B; Hahn, Daniel A

    2016-09-14

    Metabolic flexibility is an important component of adaptation to stressful environments, including thermal stress and latitudinal adaptation. A long history of population genetic studies suggest that selection on core metabolic enzymes may shape life histories by altering metabolic flux. However, the direct relationship between selection on thermal stress hardiness and metabolic flux has not previously been tested. We investigated flexibility of nutrient catabolism during cold stress in Drosophila melanogaster artificially selected for fast or slow recovery from chill coma (i.e. cold-hardy or -susceptible), specifically testing the hypothesis that stress adaptation increases metabolic turnover. Using (13)C-labelled glucose, we first showed that cold-hardy flies more rapidly incorporate ingested carbon into amino acids and newly synthesized glucose, permitting rapid synthesis of proline, a compound shown elsewhere to improve survival of cold stress. Second, using glucose and leucine tracers we showed that cold-hardy flies had higher oxidation rates than cold-susceptible flies before cold exposure, similar oxidation rates during cold exposure, and returned to higher oxidation rates during recovery. Additionally, cold-hardy flies transferred compounds among body pools more rapidly during cold exposure and recovery. Increased metabolic turnover may allow cold-adapted flies to better prepare for, resist and repair/tolerate cold damage. This work illustrates for the first time differences in nutrient fluxes associated with cold adaptation, suggesting that metabolic costs associated with cold hardiness could invoke resource-based trade-offs that shape life histories. PMID:27605506

  9. Cold adaptation increases rates of nutrient flow and metabolic plasticity during cold exposure in Drosophila melanogaster.

    PubMed

    Williams, Caroline M; McCue, Marshall D; Sunny, Nishanth E; Szejner-Sigal, Andre; Morgan, Theodore J; Allison, David B; Hahn, Daniel A

    2016-09-14

    Metabolic flexibility is an important component of adaptation to stressful environments, including thermal stress and latitudinal adaptation. A long history of population genetic studies suggest that selection on core metabolic enzymes may shape life histories by altering metabolic flux. However, the direct relationship between selection on thermal stress hardiness and metabolic flux has not previously been tested. We investigated flexibility of nutrient catabolism during cold stress in Drosophila melanogaster artificially selected for fast or slow recovery from chill coma (i.e. cold-hardy or -susceptible), specifically testing the hypothesis that stress adaptation increases metabolic turnover. Using (13)C-labelled glucose, we first showed that cold-hardy flies more rapidly incorporate ingested carbon into amino acids and newly synthesized glucose, permitting rapid synthesis of proline, a compound shown elsewhere to improve survival of cold stress. Second, using glucose and leucine tracers we showed that cold-hardy flies had higher oxidation rates than cold-susceptible flies before cold exposure, similar oxidation rates during cold exposure, and returned to higher oxidation rates during recovery. Additionally, cold-hardy flies transferred compounds among body pools more rapidly during cold exposure and recovery. Increased metabolic turnover may allow cold-adapted flies to better prepare for, resist and repair/tolerate cold damage. This work illustrates for the first time differences in nutrient fluxes associated with cold adaptation, suggesting that metabolic costs associated with cold hardiness could invoke resource-based trade-offs that shape life histories.

  10. The Antarctic Chlamydomonas raudensis: an emerging model for cold adaptation of photosynthesis.

    PubMed

    Dolhi, Jenna M; Maxwell, Denis P; Morgan-Kiss, Rachael M

    2013-09-01

    Permanently cold habitats dominate our planet and psychrophilic microorganisms thrive in cold environments. Environmental adaptations unique to psychrophilic microorganisms have been thoroughly described; however, the vast majority of studies to date have focused on cold-adapted bacteria. The combination of low temperatures in the presence of light is one of the most damaging environmental stresses for a photosynthetic organism: in order to survive, photopsychrophiles (i.e. photosynthetic organisms adapted to low temperatures) balance temperature-independent reactions of light energy capture/transduction with downstream temperature-dependent metabolic processes such as carbon fixation. Here, we review research on photopsychrophiles with a focus on an emerging model organism, Chlamydomonas raudensis UWO241 (UWO241). UWO241 is a psychrophilic green algal species and is a member of the photosynthetic microbial eukaryote community that provides the majority of fixed carbon for ice-covered lake ecosystems located in the McMurdo Dry Valleys, Antarctica. The water column exerts a range of environmental stressors on the phytoplankton community that inhabits this aquatic ecosystem, including low temperatures, extreme shade of an unusual spectral range (blue-green), high salinity, nutrient deprivation and extremes in seasonal photoperiod. More than two decades of work on UWO241 have produced one of our most comprehensive views of environmental adaptation in a cold-adapted, photosynthetic microbial eukaryote.

  11. Diversity and bioprospecting of fungal communities associated with endemic and cold-adapted macroalgae in Antarctica

    PubMed Central

    Godinho, Valéria M; Furbino, Laura E; Santiago, Iara F; Pellizzari, Franciane M; Yokoya, Nair S; Pupo, Diclá; Alves, Tânia MA; S Junior, Policarpo A; Romanha, Alvaro J; Zani, Carlos L; Cantrell, Charles L; Rosa, Carlos A; Rosa, Luiz H

    2013-01-01

    We surveyed the distribution and diversity of fungi associated with eight macroalgae from Antarctica and their capability to produce bioactive compounds. The collections yielded 148 fungal isolates, which were identified using molecular methods as belonging to 21 genera and 50 taxa. The most frequent taxa were Geomyces species (sp.), Penicillium sp. and Metschnikowia australis. Seven fungal isolates associated with the endemic Antarctic macroalgae Monostroma hariotii (Chlorophyte) displayed high internal transcribed spacer sequences similarities with the psychrophilic pathogenic fungus Geomyces destructans. Thirty-three fungal singletons (66%) were identified, representing rare components of the fungal communities. The fungal communities displayed high diversity, richness and dominance indices; however, rarefaction curves indicated that not all of the fungal diversity present was recovered. Penicillium sp. UFMGCB 6034 and Penicillium sp. UFMGCB 6120, recovered from the endemic species Palmaria decipiens (Rhodophyte) and M. hariotii, respectively, yielded extracts with high and selective antifungal and/or trypanocidal activities, in which a preliminary spectral analysis using proton nuclear magnetic resonance spectroscopy indicated the presence of highly functionalised aromatic compounds. These results suggest that the endemic and cold-adapted macroalgae of Antarctica shelter a rich, diversity and complex fungal communities consisting of a few dominant indigenous or mesophilic cold-adapted species, and a large number of rare and/or endemic taxa, which may provide an interesting model of algal–fungal interactions under extreme conditions as well as a potential source of bioactive compounds. PMID:23702515

  12. Cold adaptation in the marine bacterium, Sphingopyxis alaskensis, assessed using quantitative proteomics.

    PubMed

    Ting, Lily; Williams, Timothy J; Cowley, Mark J; Lauro, Federico M; Guilhaus, Michael; Raftery, Mark J; Cavicchioli, Ricardo

    2010-10-01

    The cold marine environment constitutes a large proportion of the Earth's biosphere. Sphingopyxis alaskensis was isolated as a numerically abundant bacterium from several cold marine locations, and has been extensively studied as a model marine bacterium. Recently, a metabolic labelling platform was developed to comprehensively identify and quantify proteins from S. alaskensis. The approach incorporated data normalization and statistical validation for the purpose of generating highly confident quantitative proteomics data. Using this approach, we determined quantitative differences between cells grown at 10°C (low temperature) and 30°C (high temperature). Cold adaptation was linked to specific aspects of gene expression: a dedicated protein-folding system using GroESL, DnaK, DnaJ, GrpE, SecB, ClpB and PPIase; polyhydroxyalkanoate-associated storage materials; a link between enzymes in fatty acid metabolism and energy generation; de novo synthesis of polyunsaturated fatty acids in the membrane and cell wall; inorganic phosphate ion transport by a phosphate import PstB homologue; TonB-dependent receptor and bacterioferritin in iron homeostasis; histidine, tryptophan and proline amino acid metabolism; and a large number of proteins without annotated functions. This study provides a new level of understanding on how important marine bacteria can adapt to compete effectively in cold marine environments. This study is also a benchmark for comparative proteomic analyses with other important marine bacteria and other cold-adapted organisms. PMID:20482592

  13. Optimization of cold-adapted alpha-galactosidase expression in Escherichia coli.

    PubMed

    Golotin, V A; Balabanova, L A; Noskova, Yu A; Slepchenko, L V; Bakunina, I Yu; Vorobieva, N S; Terenteva, N A; Rasskazov, V A

    2016-07-01

    α-Galactosidase (α-PsGal) of the cold-adapted marine bacterium Pseudoalteromonas sp. KMM 701 was cloned into the pET-40b(+) vector to study its properties and to develop an effective method for modifying human B-erythrocytes into O-blood group. The use of heat-shock as a pre-induction treatment, IPTG concentration of 0.2 mM and post-induction cultivation at 18 °C for 20 h in the developed MX-medium allowed increasing the recombinant Escherichia coli Rosetta (DE3)/40Gal strain productivity up to 30 times and the total soluble α-PsGal yield up to 40 times. PMID:27033343

  14. Cold adaptation of the mononuclear molybdoenzyme periplasmic nitrate reductase from the Antarctic bacterium Shewanella gelidimarina

    SciTech Connect

    Simpson, Philippa J.L.; Codd, Rachel

    2011-11-04

    Highlights: Black-Right-Pointing-Pointer Cold-adapted phenotype of NapA from the Antarctic bacterium Shewanella gelidimarina. Black-Right-Pointing-Pointer Protein homology model of NapA from S. gelidimarina and mesophilic homologue. Black-Right-Pointing-Pointer Six amino acid residues identified as lead candidates governing NapA cold adaptation. Black-Right-Pointing-Pointer Molecular-level understanding of designing cool-temperature in situ oxyanion sensors. -- Abstract: The reduction of nitrate to nitrite is catalysed in bacteria by periplasmic nitrate reductase (Nap) which describes a system of variable protein subunits encoded by the nap operon. Nitrate reduction occurs in the NapA subunit, which contains a bis-molybdopterin guanine dinucleotide (Mo-MGD) cofactor and one [4Fe-4S] iron-sulfur cluster. The activity of periplasmic nitrate reductase (Nap) isolated as native protein from the cold-adapted (psychrophilic) Antarctic bacterium Shewanella gelidimarina (Nap{sub Sgel}) and middle-temperature adapted (mesophilic) Shewanella putrefaciens (Nap{sub Sput}) was examined at varied temperature. Irreversible deactivation of Nap{sub Sgel} and Nap{sub Sput} occurred at 54.5 and 65 Degree-Sign C, respectively. When Nap{sub Sgel} was preincubated at 21-70 Degree-Sign C for 30 min, the room-temperature nitrate reductase activity was maximal and invariant between 21 and 54 Degree-Sign C, which suggested that Nap{sub Sgel} was poised for optimal catalysis at modest temperatures and, unlike Nap{sub Sput}, did not benefit from thermally-induced refolding. At 20 Degree-Sign C, Nap{sub Sgel} reduced selenate at 16% of the rate of nitrate reduction. Nap{sub Sput} did not reduce selenate. Sequence alignment showed 46 amino acid residue substitutions in Nap{sub Sgel} that were conserved in NapA from mesophilic Shewanella, Rhodobacter and Escherichia species and could be associated with the Nap{sub Sgel} cold-adapted phenotype. Protein homology modeling of Nap{sub Sgel} using a

  15. Flexibility and Stability Trade-Off in Active Site of Cold-Adapted Pseudomonas mandelii Esterase EstK.

    PubMed

    Truongvan, Ngoc; Jang, Sei-Heon; Lee, ChangWoo

    2016-06-28

    Cold-adapted enzymes exhibit enhanced conformational flexibility, especially in their active sites, as compared with their warmer-temperature counterparts. However, the mechanism by which cold-adapted enzymes maintain their active site stability is largely unknown. In this study, we investigated the role of conserved D308-Y309 residues located in the same loop as the catalytic H307 residue in the cold-adapted esterase EstK from Pseudomonas mandelii. Mutation of D308 and/or Y309 to Ala or deletion resulted in increased conformational flexibility. Particularly, the D308A or Y309A mutant showed enhanced substrate affinity and catalytic rate, as compared with wild-type EstK, via enlargement of the active site. However, all mutant EstK enzymes exhibited reduced thermal stability. The effect of mutation was greater for D308 than Y309. These results indicate that D308 is not preferable for substrate selection and catalytic activity, whereas hydrogen bond formation involving D308 is critical for active site stabilization. Taken together, conformation of the EstK active site is constrained via flexibility-stability trade-off for enzyme catalysis and thermal stability. Our study provides further insights into active site stabilization of cold-adapted enzymes. PMID:27259687

  16. Cloning, expression and structural stability of a cold-adapted ß-Galactosidase from Rahnella sp.R3

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A novel gene was isolated for the first time from a psychrophilic gram-negative bacterium Rahnella sp.R3. It encoded a cold-adapted ß-galactosidase (R-ß-Gal). Recombinant R-ß-Gal was expressed in Escherichia coli BL21 (DE3), purified, and characterized. R-ß-Gal belongs to the glycosyl hydrolase fami...

  17. Hot experience for cold-adapted microorganisms: temperature sensitivity of soil enzymes

    NASA Astrophysics Data System (ADS)

    Liu, Shibin; Razavidezfuly, Baharsadat; Kuzyakov, Yakov

    2016-04-01

    The temperature sensitivity of enzymes responsible for organic matter decomposition in cold environment soil, where warming is expected to be greatest is crucial. Based on Michaelis-Menten kinetics and Arrhenius function, we hypothesized that cold-adapted microorganisms will produce high efficient enzymes at cold temperatures (enzymes with lower apparent activation energy (Ea) at cold temperature ranges). To test our hypothesis, 30 g soil of Tibetan Plateau (4100 m a.s.l., annual temperature 2.4 °C) in 4 replicates were incubated for one month over a temperature range of 0-40 °C (with 5 °C steps) and determined the kinetic parameters of six enzymes involved in decomposing organics: cellobiohydrolase and β-glucosidase, which are commonly measured as enzymes responsible for consecutive stages of cellulose degradation; xylanase, which is responsible for breaking down hemicelluloses; acid phosphatase, which mineralizes organic P to phosphate by hydrolyzing phosphoric (mono) ester bonds under acidic conditions. Activities of leucine aminopeptidase and tyrosine aminopeptidase were analyzed to assess the hydrolysis of L-peptide bonds. The apparent activation energy varied between enzymes from 42 (phosphatase) to 54 (cellobiohydrolase) kJ mol-1 corresponding to the Q10 values of the enzyme reactions of 1.8-2.3. The increase of substrate affinity (Km) with temperature was gradual for most tested enzymes from 0-20 °C (enzymes involved in C cycle), (proteases) and 0-40 °C (phosphatase). However, within a high range of temperatures (25-40 °C) the hydrolytic activity was governed by enzymes with nearly constant substrate affinity. Overall, for enzymes involved in C cycle and proteases, a strong increase (30-40%) in Km at high temperatures (25 °C) reflects an expression of multiple isoenzymes each with different temperature optima and probable shift of microbial community. The general trend of catalytic efficiency (Vmax/Km) demonstrated a gradual increase with

  18. Cold adaptation of the mononuclear molybdoenzyme periplasmic nitrate reductase from the Antarctic bacterium Shewanella gelidimarina.

    PubMed

    Simpson, Philippa J L; Codd, Rachel

    2011-11-01

    The reduction of nitrate to nitrite is catalysed in bacteria by periplasmic nitrate reductase (Nap) which describes a system of variable protein subunits encoded by the nap operon. Nitrate reduction occurs in the NapA subunit, which contains a bis-molybdopterin guanine dinucleotide (Mo-MGD) cofactor and one [4Fe-4S] iron-sulfur cluster. The activity of periplasmic nitrate reductase (Nap) isolated as native protein from the cold-adapted (psychrophilic) Antarctic bacterium Shewanella gelidimarina (Nap(Sgel)) and middle-temperature adapted (mesophilic) Shewanella putrefaciens (Nap(Sput)) was examined at varied temperature. Irreversible deactivation of Nap(Sgel) and Nap(Sput) occurred at 54.5 and 65°C, respectively. When Nap(Sgel) was preincubated at 21-70°C for 30 min, the room-temperature nitrate reductase activity was maximal and invariant between 21 and 54°C, which suggested that Nap(Sgel) was poised for optimal catalysis at modest temperatures and, unlike Nap(Sput), did not benefit from thermally-induced refolding. At 20°C, Nap(Sgel) reduced selenate at 16% of the rate of nitrate reduction. Nap(Sput) did not reduce selenate. Sequence alignment showed 46 amino acid residue substitutions in Nap(Sgel) that were conserved in NapA from mesophilic Shewanella, Rhodobacter and Escherichia species and could be associated with the Nap(Sgel) cold-adapted phenotype. Protein homology modeling of Nap(Sgel) using a mesophilic template with 66% amino acid identity showed the majority of substitutions occurred at the protein surface distal to the Mo-MGD cofactor. Two mesophilic↔psychrophilic substitutions (Asn↔His, Val↔Trp) occurred in a region close to the surface of the NapA substrate funnel resulting in potential interdomain π-π and/or cation-π interactions. Three mesophilic↔psychrophilic substitutions occurred within 4.5Å of the Mo-MGD cofactor (Phe↔Met, Ala↔Ser, Ser↔Thr) resulting in local regions that varied in hydrophobicity and hydrogen bonding

  19. Cold adaptation shapes the robustness of metabolic networks in Drosophila melanogaster.

    PubMed

    Williams, Caroline M; Watanabe, Miki; Guarracino, Mario R; Ferraro, Maria B; Edison, Arthur S; Morgan, Theodore J; Boroujerdi, Arezue F B; Hahn, Daniel A

    2014-12-01

    When ectotherms are exposed to low temperatures, they enter a cold-induced coma (chill coma) that prevents resource acquisition, mating, oviposition, and escape from predation. There is substantial variation in time taken to recover from chill coma both within and among species, and this variation is correlated with habitat temperatures such that insects from cold environments recover more quickly. This suggests an adaptive response, but the mechanisms underlying variation in recovery times are unknown, making it difficult to decisively test adaptive hypotheses. We use replicated lines of Drosophila melanogaster selected in the laboratory for fast (hardy) or slow (susceptible) chill-coma recovery times to investigate modifications to metabolic profiles associated with cold adaptation. We measured metabolite concentrations of flies before, during, and after cold exposure using nuclear magnetic resonance (NMR) spectroscopy to test the hypotheses that hardy flies maintain metabolic homeostasis better during cold exposure and recovery, and that their metabolic networks are more robust to cold-induced perturbations. The metabolites of cold-hardy flies were less cold responsive and their metabolic networks during cold exposure were more robust, supporting our hypotheses. Metabolites involved in membrane lipid synthesis, tryptophan metabolism, oxidative stress, energy balance, and proline metabolism were altered by selection on cold tolerance. We discuss the potential significance of these alterations.

  20. A cold-adapted, solvent and salt tolerant esterase from marine bacterium Psychrobacter pacificensis.

    PubMed

    Wu, Gaobing; Zhang, Xiangnan; Wei, Lu; Wu, Guojie; Kumar, Ashok; Mao, Tao; Liu, Ziduo

    2015-11-01

    Lipolytic enzymes with unique physico-chemical characteristics are gaining more attention for their immense industrial importance. In this study, a novel lipolytic enzyme (Est11) was cloned from the genomic library of a marine bacterium Psychrobacter pacificensis. The enzyme was expressed in Escherichia coli and purified to homogeneity with molecular mass of 32.9kDa. The recombinant Est11 was able to hydrolyze short chain esters (C2-C8) and displayed an optimum activity against butyrate ester (C4). The optimal temperature and pH were 25°C and 7.5, respectively. Est11 retained more than 70% of its original activity at 10°C, suggesting that it was a cold-active esterase. The enzyme was highly active and stable at high concentration of NaCl (5M). Further, incubation with ethanol, isopropanol, propanediol, DMSO, acetonitrile, and glycerol rendered remarkable positive effects on Est11 activity. Typically, even at the concentration of 30% (v/v), ethanol, DMSO, and propanediol increased Est11 activity by 1.3, 2.0, and 2.4-folds, respectively. This new robust enzyme with remarkable properties like cold-adaptability, exceptional tolerance to salt and organic solvents provides us a promising candidate to meet the needs of some harsh industrial processes. PMID:26231332

  1. Expression and purification of a cold-adapted group III trypsin in Escherichia coli.

    PubMed

    Pálsdóttir, Helga Margrét; Gudmundsdóttir, Agústa

    2007-02-01

    The recently classified group III trypsins include members like Atlantic cod (Gadus morhua) trypsin Y as well as seven analogues from other cold-adapted fish species. The eight group III trypsins have been characterized from their cDNAs and deduced amino acid sequences but none of the enzymes have been isolated from their native sources. This study describes the successful expression and purification of a recombinant HP-thioredoxin-trypsin Y fusion protein in the His-Patch ThioFusion Escherichia coli expression system and its purification by chromatographic methods. The recombinant form of trypsin Y was previously expressed in Pichia pastoris making it the first biochemically characterized group III trypsin. It has dual substrate specificity towards trypsin and chymotrypsin substrates and demonstrates an increasing activity at temperatures between 2 and 21 degrees C with a complete inactivation at 30 degrees C. The aim of the study was to facilitate further studies of recombinant trypsin Y by finding an expression system yielding higher amounts of the enzyme than possible in our hands in the P. pastoris system. Also, commercial production of trypsin Y will require an efficient and inexpensive expression system like the His-Patch ThioFusion E. coli expression system described here as the enzyme is produced in very low amounts in the Atlantic cod.

  2. Reciprocal Influence of Protein Domains in the Cold-Adapted Acyl Aminoacyl Peptidase from Sporosarcina psychrophila

    PubMed Central

    Parravicini, Federica; Natalello, Antonino; Papaleo, Elena; De Gioia, Luca; Doglia, Silvia Maria; Lotti, Marina; Brocca, Stefania

    2013-01-01

    Acyl aminoacyl peptidases are two-domain proteins composed by a C-terminal catalytic α/β-hydrolase domain and by an N-terminal β-propeller domain connected through a structural element that is at the N-terminus in sequence but participates in the 3D structure of the C-domain. We investigated about the structural and functional interplay between the two domains and the bridge structure (in this case a single helix named α1-helix) in the cold-adapted enzyme from Sporosarcina psychrophila (SpAAP) using both protein variants in which entire domains were deleted and proteins carrying substitutions in the α1-helix. We found that in this enzyme the inter-domain connection dramatically affects the stability of both the whole enzyme and the β-propeller. The α1-helix is required for the stability of the intact protein, as in other enzymes of the same family; however in this psychrophilic enzyme only, it destabilizes the isolated β-propeller. A single charged residue (E10) in the α1-helix plays a major role for the stability of the whole structure. Overall, a strict interaction of the SpAAP domains seems to be mandatory for the preservation of their reciprocal structural integrity and may witness their co-evolution. PMID:23457536

  3. Purification and characterization of cold-adapted beta-agarase from an Antarctic psychrophilic strain

    PubMed Central

    Li, Jiang; Hu, Qiushi; Li, Yuquan; Xu, Yuan

    2015-01-01

    An extracellular β-agarase was purified from Pseudoalteromonas sp. NJ21, a Psychrophilic agar-degrading bacterium isolated from Antarctic Prydz Bay sediments. The purified agarase (Aga21) revealed a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, with an apparent molecular weight of 80 kDa. The optimum pH and temperature of the agarase were 8.0 and 30 °C, respectively. However, it maintained as much as 85% of the maximum activities at 10 °C. Significant activation of the agarase was observed in the presence of Mg2+, Mn2+, K+; Ca2+, Na+, Ba2+, Zn2+, Cu2+, Co2+, Fe2+, Sr2+ and EDTA inhibited the enzyme activity. The enzymatic hydrolyzed product of agar was characterized as neoagarobiose. Furthermore, this work is the first evidence of cold-adapted agarase in Antarctic psychrophilic bacteria and these results indicate the potential for the Antarctic agarase as a catalyst in medicine, food and cosmetic industries. PMID:26413048

  4. Are mountain habitats becoming more suitable for generalist than cold-adapted lizards thermoregulation?

    PubMed

    Ortega, Zaida; Mencía, Abraham; Pérez-Mellado, Valentín

    2016-01-01

    Mountain lizards are highly vulnerable to climate change, and the continuous warming of their habitats could be seriously threatening their survival. We aim to compare the thermal ecology and microhabitat selection of a mountain lizard, Iberolacerta galani, and a widely distributed lizard, Podarcis bocagei, in a montane area. Both species are currently in close syntopy in the study area, at 1,400 m above the sea level. We determined the precision, accuracy and effectiveness of thermoregulation, and the thermal quality of habitat for both species. We also compared the selection of thermal microhabitats between both species. Results show that I. galani is a cold-adapted thermal specialist with a preferred temperature range of 27.9-29.7 °C, while P. bocagei would be a thermal generalist, with a broader and higher preferred temperature range (30.1-34.5 °C). In addition, I. galani selects rocky substrates while P. bocagei selects warmer soil and leaf litter substrates. The thermal quality of the habitat is higher for P. bocagei than for I. galani. Finally, P. bocagei achieves a significantly higher effectiveness of thermoregulation (0.87) than I. galani (0.80). Therefore, these mountain habitat conditions seem currently more suitable for performance of thermophilic generalist lizards than for cold-specialist lizards.

  5. Purification and Characterization of a Cold-Adapted Lipase from Oceanobacillus Strain PT-11

    PubMed Central

    Jiewei, Tian; Zuchao, Lei; Peng, Qiu; Lei, Wang; Yongqiang, Tian

    2014-01-01

    We isolated a moderately halophilic lipase-producing bacterium from the saline soil. Based on the morphological, physiological, chemotaxonomic and phylogenetic analysis, the isolate PT-11 was postulated to be a novel species identified as Oceanobacillus rekensis PT-11. The lipase was purified 2.50-fold by Q-Sepharose FF and SP-Sepharose FF chromatography and its molecular mass was estimated to be 23.5 kDa by SDS-PAGE. It was highly active over the broad temperature ranging from 10 to 35°C and showed up to 80% of the maximum activity at 10°C indicating the lipase to be a typical cold-adapted enzyme. The enzyme activity was slightly enhanced by Na+, Li+ and K+. Incubation with detergents, such as Tween-20 and Tween-80, slightly inhibited the enzyme activity; while Triton X-100decreased the enzyme activity. The enzyme was fairly stable in the presence of long-chain alcohols but was highly denatured in hydrophilic solvents such as acetone or short-chain alcohols (C1–C3). PMID:24984141

  6. Cold adaptation shapes the robustness of metabolic networks in Drosophila melanogaster

    PubMed Central

    Williams, CM; Watanabe, M; Guarracino, MR; Ferraro, MB; Edison, AS; Morgan, TJ; Boroujerdi, AFB; Hahn, DA

    2015-01-01

    When ectotherms are exposed to low temperatures, they enter a cold-induced coma (chill coma) that prevents resource acquisition, mating, oviposition, and escape from predation. There is substantial variation in time taken to recover from chill coma both within and among species, and this variation is correlated with habitat temperatures such that insects from cold environments recover more quickly. This suggests an adaptive response, but the mechanisms underlying variation in recovery times are unknown, making it difficult to decisively test adaptive hypotheses. We use replicated lines of Drosophila melanogaster selected in the laboratory for fast (hardy) or slow (susceptible) chill-coma recovery times to investigate modifications to metabolic profiles associated with cold adaptation. We measured metabolite concentrations of flies before, during, and after cold exposure using NMR spectroscopy to test the hypotheses that hardy flies maintain metabolic homeostasis better during cold exposure and recovery, and that their metabolic networks are more robust to cold-induced perturbations. The metabolites of cold-hardy flies were less cold responsive and their metabolic networks during cold exposure were more robust, supporting our hypotheses. Metabolites involved in membrane lipid synthesis, tryptophan metabolism, oxidative stress, energy balance, and proline metabolism were altered by selection on cold tolerance. We discuss the potential significance of these alterations. PMID:25308124

  7. Structural characterization of metal binding to a cold-adapted frataxin.

    PubMed

    Noguera, Martín E; Roman, Ernesto A; Rigal, Juan B; Cousido-Siah, Alexandra; Mitschler, André; Podjarny, Alberto; Santos, Javier

    2015-06-01

    Frataxin is an evolutionary conserved protein that participates in iron metabolism. Deficiency of this small protein in humans causes a severe neurodegenerative disease known as Friedreich's ataxia. A number of studies indicate that frataxin binds iron and regulates Fe-S cluster biosynthesis. Previous structural studies showed that metal binding occurs mainly in a region of high density of negative charge. However, a comprehensive characterization of the binding sites is required to gain further insights into the mechanistic details of frataxin function. In this work, we have solved the X-ray crystal structures of a cold-adapted frataxin from a psychrophilic bacterium in the presence of cobalt or europium ions. We have identified a number of metal-binding sites, mainly solvent exposed, several of which had not been observed in previous studies on mesophilic homologues. No major structural changes were detected upon metal binding, although the structures exhibit significant changes in crystallographic B-factors. The analysis of these B-factors, in combination with crystal packing and RMSD among structures, suggests the existence of localized changes in the internal motions. Based on these results, we propose that bacterial frataxins possess binding sites of moderate affinity for a quick capture and transfer of iron to other proteins and for the regulation of Fe-S cluster biosynthesis, modulating interactions with partner proteins.

  8. Purification and characterization of a cold-adapted lipase from Oceanobacillus strain PT-11.

    PubMed

    Jiewei, Tian; Zuchao, Lei; Peng, Qiu; Lei, Wang; Yongqiang, Tian

    2014-01-01

    We isolated a moderately halophilic lipase-producing bacterium from the saline soil. Based on the morphological, physiological, chemotaxonomic and phylogenetic analysis, the isolate PT-11 was postulated to be a novel species identified as Oceanobacillus rekensis PT-11. The lipase was purified 2.50-fold by Q-Sepharose FF and SP-Sepharose FF chromatography and its molecular mass was estimated to be 23.5 kDa by SDS-PAGE. It was highly active over the broad temperature ranging from 10 to 35°C and showed up to 80% of the maximum activity at 10°C indicating the lipase to be a typical cold-adapted enzyme. The enzyme activity was slightly enhanced by Na+, Li+ and K+. Incubation with detergents, such as Tween-20 and Tween-80, slightly inhibited the enzyme activity; while Triton X-100decreased the enzyme activity. The enzyme was fairly stable in the presence of long-chain alcohols but was highly denatured in hydrophilic solvents such as acetone or short-chain alcohols (C1-C3). PMID:24984141

  9. Are mountain habitats becoming more suitable for generalist than cold-adapted lizards thermoregulation?

    PubMed Central

    Mencía, Abraham; Pérez-Mellado, Valentín

    2016-01-01

    Mountain lizards are highly vulnerable to climate change, and the continuous warming of their habitats could be seriously threatening their survival. We aim to compare the thermal ecology and microhabitat selection of a mountain lizard, Iberolacerta galani, and a widely distributed lizard, Podarcis bocagei, in a montane area. Both species are currently in close syntopy in the study area, at 1,400 m above the sea level. We determined the precision, accuracy and effectiveness of thermoregulation, and the thermal quality of habitat for both species. We also compared the selection of thermal microhabitats between both species. Results show that I. galani is a cold-adapted thermal specialist with a preferred temperature range of 27.9–29.7 °C, while P. bocagei would be a thermal generalist, with a broader and higher preferred temperature range (30.1–34.5 °C). In addition, I. galani selects rocky substrates while P. bocagei selects warmer soil and leaf litter substrates. The thermal quality of the habitat is higher for P. bocagei than for I. galani. Finally, P. bocagei achieves a significantly higher effectiveness of thermoregulation (0.87) than I. galani (0.80). Therefore, these mountain habitat conditions seem currently more suitable for performance of thermophilic generalist lizards than for cold-specialist lizards. PMID:27280076

  10. Duration of immunity engendered by a single dose of a cold-adapted strain of Avian pneumovirus

    PubMed Central

    2006-01-01

    Abstract The duration of immunity after a single dose of a cold-adapted strain of Avian pneumovirus (APV) was studied. Turkeys were vaccinated at 1 wk of age and challenged with virulent virus 3, 7, 10, and 14 wk later. Nonvaccinated groups were also challenged at the same times. No clinical signs were observed in the vaccinated birds after vaccination or after any challenge. No viral RNA was shed by the vaccinated birds after any challenge. The nonvaccinated birds shed viral RNA after all challenges. Avian pneumovirus-specific humoral antibodies were detected in the vaccinated birds until 14 wk after vaccination. The results of this preliminary study indicate that inoculation with a single dose of a cold-adapted strain of APV at 1 wk of age provides protection until 15 wk of age. PMID:16548335

  11. Aerobic and Anaerobic Thiosulfate Oxidation by a Cold-Adapted, Subglacial Chemoautotroph

    PubMed Central

    Harrold, Zoë R.; Skidmore, Mark L.; Hamilton, Trinity L.; Desch, Libby; Amada, Kirina; van Gelder, Will; Glover, Kevin; Roden, Eric E.

    2015-01-01

    Geochemical data indicate that protons released during pyrite (FeS2) oxidation are important drivers of mineral weathering in oxic and anoxic zones of many aquatic environments, including those beneath glaciers. Oxidation of FeS2 under oxic, circumneutral conditions proceeds through the metastable intermediate thiosulfate (S2O32−), which represents an electron donor capable of supporting microbial metabolism. Subglacial meltwaters sampled from Robertson Glacier (RG), Canada, over a seasonal melt cycle revealed concentrations of S2O32− that were typically below the limit of detection, despite the presence of available pyrite and concentrations of the FeS2 oxidation product sulfate (SO42−) several orders of magnitude higher than those of S2O32−. Here we report on the physiological and genomic characterization of the chemolithoautotrophic facultative anaerobe Thiobacillus sp. strain RG5 isolated from the subglacial environment at RG. The RG5 genome encodes genes involved with pathways for the complete oxidation of S2O32−, CO2 fixation, and aerobic and anaerobic respiration with nitrite or nitrate. Growth experiments indicated that the energy required to synthesize a cell under oxygen- or nitrate-reducing conditions with S2O32− as the electron donor was lower at 5.1°C than 14.4°C, indicating that this organism is cold adapted. RG sediment-associated transcripts of soxB, which encodes a component of the S2O32−-oxidizing complex, were closely affiliated with soxB from RG5. Collectively, these results suggest an active sulfur cycle in the subglacial environment at RG mediated in part by populations closely affiliated with RG5. The consumption of S2O32− by RG5-like populations may accelerate abiotic FeS2 oxidation, thereby enhancing mineral weathering in the subglacial environment. PMID:26712544

  12. Genome-wide analysis of cold adaptation in indigenous Siberian populations.

    PubMed

    Cardona, Alexia; Pagani, Luca; Antao, Tiago; Lawson, Daniel J; Eichstaedt, Christina A; Yngvadottir, Bryndis; Shwe, Ma Than Than; Wee, Joseph; Romero, Irene Gallego; Raj, Srilakshmi; Metspalu, Mait; Villems, Richard; Willerslev, Eske; Tyler-Smith, Chris; Malyarchuk, Boris A; Derenko, Miroslava V; Kivisild, Toomas

    2014-01-01

    Following the dispersal out of Africa, where hominins evolved in warm environments for millions of years, our species has colonised different climate zones of the world, including high latitudes and cold environments. The extent to which human habitation in (sub-)Arctic regions has been enabled by cultural buffering, short-term acclimatization and genetic adaptations is not clearly understood. Present day indigenous populations of Siberia show a number of phenotypic features, such as increased basal metabolic rate, low serum lipid levels and increased blood pressure that have been attributed to adaptation to the extreme cold climate. In this study we introduce a dataset of 200 individuals from ten indigenous Siberian populations that were genotyped for 730,525 SNPs across the genome to identify genes and non-coding regions that have undergone unusually rapid allele frequency and long-range haplotype homozygosity change in the recent past. At least three distinct population clusters could be identified among the Siberians, each of which showed a number of unique signals of selection. A region on chromosome 11 (chr11:66-69 Mb) contained the largest amount of clustering of significant signals and also the strongest signals in all the different selection tests performed. We present a list of candidate cold adaption genes that showed significant signals of positive selection with our strongest signals associated with genes involved in energy regulation and metabolism (CPT1A, LRP5, THADA) and vascular smooth muscle contraction (PRKG1). By employing a new method that paints phased chromosome chunks by their ancestry we distinguish local Siberian-specific long-range haplotype signals from those introduced by admixture. PMID:24847810

  13. Cold adaptation mechanisms in the ghost moth Hepialus xiaojinensis: Metabolic regulation and thermal compensation.

    PubMed

    Zhu, Wei; Zhang, Huan; Li, Xuan; Meng, Qian; Shu, Ruihao; Wang, Menglong; Zhou, Guiling; Wang, Hongtuo; Miao, Lin; Zhang, Jihong; Qin, Qilian

    2016-02-01

    Ghost moths (Lepidoptera: Hepialidae) are cold-adapted stenothermal species inhabiting alpine meadows on the Tibetan Plateau. They have an optimal developmental temperature of 12-16 °C but can maintain feeding and growth at 0 °C. Their survival strategies have received little attention, but these insects are a promising model for environmental adaptation. Here, biochemical adaptations and energy metabolism in response to cold were investigated in larvae of the ghost moth Hepialus xiaojinensis. Metabolic rate and respiratory quotient decreased dramatically with decreasing temperature (15-4 °C), suggesting that the energy metabolism of ghost moths, especially glycometabolism, was sensitive to cold. However, the metabolic rate at 4 °C increased with the duration of cold exposure, indicating thermal compensation to sustain energy budgets under cold conditions. Underlying regulation strategies were studied by analyzing metabolic differences between cold-acclimated (4 °C for 48 h) and control larvae (15 °C). In cold-acclimated larvae, the energy generating pathways of carbohydrates, instead of the overall consumption of carbohydrates, was compensated in the fat body by improving the transcription of related enzymes. The mobilization of lipids was also promoted, with higher diacylglycerol, monoacylglycerol and free fatty acid content in hemolymph. These results indicated that cold acclimation induced a reorganization on metabolic structure to prioritise energy metabolism. Within the aerobic process, flux throughout the tricarboxylic acid (TCA) cycle was encouraged in the fat body, and the activity of α-ketoglutarate dehydrogenase was the likely compensation target. Increased mitochondrial cristae density was observed in the midgut of cold-acclimated larvae. The thermal compensation strategies in this ghost moth span the entire process of energy metabolism, including degration of metabolic substrate, TCA cycle and oxidative phosphorylation, and from an energy budget

  14. Aerobic and Anaerobic Thiosulfate Oxidation by a Cold-Adapted, Subglacial Chemoautotroph.

    PubMed

    Harrold, Zoë R; Skidmore, Mark L; Hamilton, Trinity L; Desch, Libby; Amada, Kirina; van Gelder, Will; Glover, Kevin; Roden, Eric E; Boyd, Eric S

    2016-03-01

    Geochemical data indicate that protons released during pyrite (FeS2) oxidation are important drivers of mineral weathering in oxic and anoxic zones of many aquatic environments, including those beneath glaciers. Oxidation of FeS2 under oxic, circumneutral conditions proceeds through the metastable intermediate thiosulfate (S2O3 (2-)), which represents an electron donor capable of supporting microbial metabolism. Subglacial meltwaters sampled from Robertson Glacier (RG), Canada, over a seasonal melt cycle revealed concentrations of S2O3 (2-) that were typically below the limit of detection, despite the presence of available pyrite and concentrations of the FeS2 oxidation product sulfate (SO4 (2-)) several orders of magnitude higher than those of S2O3 (2-). Here we report on the physiological and genomic characterization of the chemolithoautotrophic facultative anaerobe Thiobacillus sp. strain RG5 isolated from the subglacial environment at RG. The RG5 genome encodes genes involved with pathways for the complete oxidation of S2O3 (2-), CO2 fixation, and aerobic and anaerobic respiration with nitrite or nitrate. Growth experiments indicated that the energy required to synthesize a cell under oxygen- or nitrate-reducing conditions with S2O3 (2-) as the electron donor was lower at 5.1°C than 14.4°C, indicating that this organism is cold adapted. RG sediment-associated transcripts of soxB, which encodes a component of the S2O3 (2-)-oxidizing complex, were closely affiliated with soxB from RG5. Collectively, these results suggest an active sulfur cycle in the subglacial environment at RG mediated in part by populations closely affiliated with RG5. The consumption of S2O3 (2-) by RG5-like populations may accelerate abiotic FeS2 oxidation, thereby enhancing mineral weathering in the subglacial environment. PMID:26712544

  15. Aerobic and Anaerobic Thiosulfate Oxidation by a Cold-Adapted, Subglacial Chemoautotroph.

    PubMed

    Harrold, Zoë R; Skidmore, Mark L; Hamilton, Trinity L; Desch, Libby; Amada, Kirina; van Gelder, Will; Glover, Kevin; Roden, Eric E; Boyd, Eric S

    2015-12-28

    Geochemical data indicate that protons released during pyrite (FeS2) oxidation are important drivers of mineral weathering in oxic and anoxic zones of many aquatic environments, including those beneath glaciers. Oxidation of FeS2 under oxic, circumneutral conditions proceeds through the metastable intermediate thiosulfate (S2O3 (2-)), which represents an electron donor capable of supporting microbial metabolism. Subglacial meltwaters sampled from Robertson Glacier (RG), Canada, over a seasonal melt cycle revealed concentrations of S2O3 (2-) that were typically below the limit of detection, despite the presence of available pyrite and concentrations of the FeS2 oxidation product sulfate (SO4 (2-)) several orders of magnitude higher than those of S2O3 (2-). Here we report on the physiological and genomic characterization of the chemolithoautotrophic facultative anaerobe Thiobacillus sp. strain RG5 isolated from the subglacial environment at RG. The RG5 genome encodes genes involved with pathways for the complete oxidation of S2O3 (2-), CO2 fixation, and aerobic and anaerobic respiration with nitrite or nitrate. Growth experiments indicated that the energy required to synthesize a cell under oxygen- or nitrate-reducing conditions with S2O3 (2-) as the electron donor was lower at 5.1°C than 14.4°C, indicating that this organism is cold adapted. RG sediment-associated transcripts of soxB, which encodes a component of the S2O3 (2-)-oxidizing complex, were closely affiliated with soxB from RG5. Collectively, these results suggest an active sulfur cycle in the subglacial environment at RG mediated in part by populations closely affiliated with RG5. The consumption of S2O3 (2-) by RG5-like populations may accelerate abiotic FeS2 oxidation, thereby enhancing mineral weathering in the subglacial environment.

  16. Diversity and cold adaptation of microorganisms isolated from the Schirmacher Oasis, Antarctica

    NASA Astrophysics Data System (ADS)

    Mojib, Nazia; Bej, Asim K.; Hoover, Richard

    2008-08-01

    We have investigated the feasibility of the PCR amplification of the 16S rRNA genes from eubacteria and Archea on samples collected on Whatman FTA filters from Schirmacher Oasis for the study of culture-independent analysis of the microbial diversity. Both conventional PCR and real-time TaqmaTM PCR successfully amplified the targeted genes. A number of diverse groups of psychrotolerant microorganisms with various pigments have been isolated when cultured on agar medium. 16S rRNA gene analysis of these isolates helped us to identify closest taxonomic genus Pseudomonas, Frigoribacterium, Arthrobacter, Flavobacterium, and Janthinobacterium. It is possible that the pigments play protective role from solar UV radiation, which is prevalent in Antarctic continent especially during Austral summer months. Study of the expression of cold adaptive protein CapB and ice-binding protein IBP using western blots showed positive detection of both or either of these proteins in 6 out of 8 isolates. Since the CapB and IBP protein structure greatly varies in microorganisms, it is possible that the 2 isolates with negative results could have a different class of these proteins. The expression of the CapB and the IBP in these isolates suggest that these proteins are essential for the survival in the Antarctic cold and subzero temperatures and protect themselves from freeze-damage. The current study provided sufficient data to further investigate the rich and diverse biota of psychrotolerant extremophiles in the Antarctic Schirmacher Oasis using both culture-independent and culture-based approaches; and understand the mechanisms of cold tolerance.

  17. Novel cold-adaptive Penicillium strain FS010 secreting thermo-labile xylanase isolated from Yellow Sea.

    PubMed

    Hou, Yun-Hua; Wang, Tian-Hong; Long, Hao; Zhu, Hui-Yuan

    2006-02-01

    A novel cold-adaptive xylanolytic Penicillium strain FS010 was isolated from Yellow sea sediments. The marine fungus grew well from 4 to 20 degrees; a lower (0 degrees) or higher (37 degrees) temperature limits its growth. The strain was identified as Penicillium chrysogenum. Compared with mesophilic P. chrysogenum, the cold-adaptive fungus secreted the cold-active xylanase (XYL) showing high hydrolytic activities at low temperature (2-15 degrees) and high sensitivity to high temperature (>50 degrees). The XYL gene was isolated from the cold-adaptive P. chrysogenum FS010 and designated as xyl. The deduced amino acid sequence of the protein encoded by xyl showed high homology with the sequence of glycoside hydrolase family 10. The gene was subcloned into an expression vector pGEX-4T-1 and the encoded protein was overexpressed as a fusion protein with glutathione-S-transferase in Escherichia coli BL21. The expression product was purified and subjected to enzymatic characterization. The optimal temperature and pH for recombinant XYL was 25 degrees and 5.5, respectively. Recombinant XYL showed nearly 80% of its maximal activity at 4 degrees and was active in the pH range 3.0-9.5.

  18. A novel cold-adapted phospholipase A(1) from Serratia sp. xjF1: Gene cloning, expression and characterization.

    PubMed

    Fu, Jianhong; Huang, Huoqing; Meng, Kun; Yuan, Tiezheng; Yao, Bin; Shi, Yuhu; Ouyang, Pingkai

    2008-01-01

    The gene encoding a cold-adapted phospholipase A(1) (PLA(1)) from a psychrotrophic, glacier soil bacterium Serratia sp. xjF1 was cloned by two-step PCR (general PCR and TAIL-PCR). The full-length fragment comprised two open reading frames plA and plS. The gene product of plA encoding 320 amino acids with a molecular weight of 33.8kDa was identified as a phospholipase A(1). Its amino acid sequence exhibited the highest homology to PLA(1) of Serratia marcescens (71%). plS encoded a protein of 251 amino acids, which showed no enzymatic activity. The result of plA expression in Escherichia coli indicated that plS might improve the efficient expression of PLA(1) in E. coli. Furthermore, PLA(1) was functionally expressed in Pichia pastoris, yielding 41.8U/mL in a 3.7L fermentor. The purified recombinant phospholipase A(1) (rPLA(1)) had features typical of cold-adapted enzymes with a temperature optimum of 35°C and a maximum activity of 70% at 10°C. The rate of catalysis was optimal at pH 9.0 and the enzyme could be slightly activated by Ca(2+). This is the first report on gene isolation and expression of cold-adapted PLA(1).

  19. Molecular determinants of enzyme cold adaptation: comparative structural and computational studies of cold- and warm-adapted enzymes.

    PubMed

    Papaleo, Elena; Tiberti, Matteo; Invernizzi, Gaetano; Pasi, Marco; Ranzani, Valeria

    2011-11-01

    The identification of molecular mechanisms underlying enzyme cold adaptation is a hot-topic both for fundamental research and industrial applications. In the present contribution, we review the last decades of structural computational investigations on cold-adapted enzymes in comparison to their warm-adapted counterparts. Comparative sequence and structural studies allow the definition of a multitude of adaptation strategies. Different enzymes carried out diverse mechanisms to adapt to low temperatures, so that a general theory for enzyme cold adaptation cannot be formulated. However, some common features can be traced in dynamic and flexibility properties of these enzymes, as well as in their intra- and inter-molecular interaction networks. Interestingly, the current data suggest that a family-centered point of view is necessary in the comparative analyses of cold- and warm-adapted enzymes. In fact, enzymes belonging to the same family or superfamily, thus sharing at least the three-dimensional fold and common features of the functional sites, have evolved similar structural and dynamic patterns to overcome the detrimental effects of low temperatures.

  20. Molecular determinants of enzyme cold adaptation: comparative structural and computational studies of cold- and warm-adapted enzymes.

    PubMed

    Papaleo, Elena; Tiberti, Matteo; Invernizzi, Gaetano; Pasi, Marco; Ranzani, Valeria

    2011-11-01

    The identification of molecular mechanisms underlying enzyme cold adaptation is a hot-topic both for fundamental research and industrial applications. In the present contribution, we review the last decades of structural computational investigations on cold-adapted enzymes in comparison to their warm-adapted counterparts. Comparative sequence and structural studies allow the definition of a multitude of adaptation strategies. Different enzymes carried out diverse mechanisms to adapt to low temperatures, so that a general theory for enzyme cold adaptation cannot be formulated. However, some common features can be traced in dynamic and flexibility properties of these enzymes, as well as in their intra- and inter-molecular interaction networks. Interestingly, the current data suggest that a family-centered point of view is necessary in the comparative analyses of cold- and warm-adapted enzymes. In fact, enzymes belonging to the same family or superfamily, thus sharing at least the three-dimensional fold and common features of the functional sites, have evolved similar structural and dynamic patterns to overcome the detrimental effects of low temperatures. PMID:21827423

  1. Enhanced catalytic site thermal stability of cold-adapted esterase EstK by a W208Y mutation.

    PubMed

    Boyineni, Jerusha; Kim, Junyoung; Kang, Beom Sik; Lee, ChangWoo; Jang, Sei-Heon

    2014-06-01

    Hydrophobic interactions are known to play an important role for cold-adaptation of proteins; however, the role of amino acid residue, Trp, has not been systematically investigated. The extracellular esterase, EstK, which was isolated from the cold-adapted bacterium Pseudomonas mandelii, has 5 Trp residues. In this study, the effects of Trp mutation on thermal stability, catalytic activity, and conformational change of EstK were investigated. Among the 5 Trp residues, W(208) was the most crucial in maintaining structural conformation and thermal stability of the enzyme. Surprisingly, mutation of W(208) to Tyr (W(208)Y) showed an increased catalytic site thermal stability at ambient temperatures with a 13-fold increase in the activity at 40°C compared to wild-type EstK. The structure model of W(208)Y suggested that Y(208) could form a hydrogen bond with D(308), which is located next to catalytic residue H(307), stabilizing the catalytic domain. Interestingly, Tyr was conserved in the corresponding position of hyper-thermophilic esterases EstE1 and AFEST, which are active at high temperatures. Our study provides a novel insight into the engineering of the catalytic site of cold-adapted enzymes with increased thermal stability and catalytic activity at ambient temperatures.

  2. [Characterization of cold-adapted influenza strain A/HongKong/1/68/162/35 as a potential donor of attenuation and high reproduction].

    PubMed

    Tsybalova, L M; Gorev, N E; Potapchuk, M V; Repko, I A; Korotkov, A V; Sergeeva, M V; Komissarov, A B; Pisareva, M M; Kuznetsov, V V; Grudinin, M P; Kiselev, O I

    2012-01-01

    Live and inactivated vaccines are currently produced using virus reassortants originating from various gene donors of internal proteins. Based on the pandemic virus A/Hong Kong/1/68 (H3N2), a cold-adapted thermo-sensitive strain A/Hong Kong/1/68/162/35 was generated. It is distinguished for its high reproductive capacity (9-9.5 lg EID50), and hemagglutinating activity (1:1024-1:2048). The strain has ts and ca phenotype: reproductive capacity at t = 39 degrees C is 1.0 lg EID50; at t = 26 degrees C, 8.5 lg EID50. A total of 16 mutations have emerged from comprehensive sequencing of the virus genome. Among them 10 mutations were located in the genes of polymerase complex and NP, with respective amino-acid substitutions. The stability of strain characteristics, such as attenuation to humans and high reproductive capacity, were confirmed by repeated sequencing of the genome after tenfold passing of the virus in chicken embryos. Reassortants of the strain A/Hong Kong/1/68/162/35 with the wild-type viruses have inherited useful features of donor virus. PMID:23477248

  3. The ability of a cold-adapted Rhodotorula mucilaginosa strain from Tibet to control blue mold in pear fruit.

    PubMed

    Hu, Hao; Yan, Fujie; Wilson, Charles; Shen, Qing; Zheng, Xiaodong

    2015-12-01

    Cold-adapted yeasts were isolated from soil samples collected in Tibet and evaluated as potential biocontrol agents against blue mold (Penicillium expansum) of pear fruit in cold storage. YC1, an isolate identified as Rhodotorula mucilaginosa, was found to exhibit the greatest biocontrol activity among the different isolates that were screened. A washed cell suspension of YC1 exhibited the best biocontrol activity among three different preparations that were used in the current study. A concentration of 10(8) cells/ml reduced the incidence of decay to 35 %, compared to the control where decay incidence was 100 %. A higher intracellular level of trehalose and a higher proportion of polyunsaturated acids present in YC1, was associated with increased the tolerance of this strain to low temperatures, relative to the other strains that were evaluated. The increased tolerance to low temperature allowed the YC1 strain of yeast to more effectively compete for nutrients and space in wounded pear fruit that had been inoculated with spores of P. expansum and placed in cold storage. The present study demonstrated the ability to select cold-adapted yeasts from cold climates and use them as biocontrol agents of postharvest diseases of fruit placed in cold storage.

  4. Effects of early cold adaptation on food efficiency and dietary-induced thermogenesis in the adult rat.

    PubMed

    Davis, J R; Tagliaferro, A R; Roberts, J S; Hill, J O

    1982-07-01

    The food intakes of male and female rats (Harvard strain) adapted to cold (5 degrees C) starting at weaning were studied in the cold and during reacclimation to 25 degrees C. Following 24 hr food deprivation, the male rats regained the weight they had lost without making up for the food they had missed (increased food efficiency), while the female rats ate 75% more food than they had missed in regaining their body weights (decreased food efficiency), independent of the temperature at which the animals were housed. On a high fat diet, both cold groups gained less weight than did the two control groups. However, each female group gained significantly less weight than its same temperature male group. Dietary-induced thermogenesis was significantly enhanced in male cold adapted rats 3 months after reacclimation to the zone of thermal neutrality but not in cold adapted female rats. Sex and strain differences are discussed. These data indicate that early chronic cold exposure results in both immediate and long-term changes in food efficiency and metabolism in adult rats, but the effects vary with the sex of the animal.

  5. Cloning, expression and structural stability of a cold-adapted β-galactosidase from Rahnella sp. R3.

    PubMed

    Fan, Yuting; Hua, Xiao; Zhang, Yuzhu; Feng, Yinghui; Shen, Qiuyun; Dong, Juan; Zhao, Wei; Zhang, Wenbin; Jin, Zhengyu; Yang, Ruijin

    2015-11-01

    A novel gene was isolated for the first time from a psychrophilic gram-negative bacterium Rahnella sp. R3. The gene encoded a cold-adapted β-galactosidase (R-β-Gal). Recombinant R-β-Gal was expressed in Escherichia coli BL21 (DE3), purified and characterized. R-β-gal belongs to the glycosyl hydrolase family 42. Circular dichroism spectrometry of the structural stability of R-β-Gal with respect to temperature indicated that the secondary structures of the enzyme were stable to 45°C. In solution, the enzyme was a homo-trimer and was active at temperatures as low as 4°C. The enzyme did not require the presence of metal ions to be active, but Mg(2+), Mn(2+), and Ca(2+) enhanced its activity slightly, whereas Fe(3+), Zn(2+) and Al(3+) appeared to inactive it. The purified enzyme displayed K(m) values of 6.5 mM for ONPG and 2.2mM for lactose at 4°C. These values were lower than the corresponding K(m)s reported for other cold-adapted β-Gals.

  6. Tenderization effect of cold-adapted collagenolytic protease MCP-01 on beef meat at low temperature and its mechanism.

    PubMed

    Zhao, Guo-Yan; Zhou, Ming-Yang; Zhao, Hui-Lin; Chen, Xiu-Lan; Xie, Bin-Bin; Zhang, Xi-Ying; He, Hai-Lun; Zhou, Bai-Cheng; Zhang, Yu-Zhong

    2012-10-15

    The enzymes currently used to increase meat tenderness are all mesophilic or thermophilic proteases. This study provides insight into the tenderization effect and the mechanism of a cold-adapted collagenolytic enzyme MCP-01 on beef meat at low temperatures. MCP-01 (10 U of caseinolytic activity) reduced the meat shear force by 23% and increased the relative myofibrillar fragmentation index of the meat by 91.7% at 4 °C, and it also kept the fresh colour and moisture of the meat. Compared to the commercially used tenderizers papain and bromelain, MCP-01 showed a unique tenderization mechanism. MCP-01 had a strong selectivity for degrading collagen at 4 °C, showed a distinct digestion pattern on the myofibrillar proteins, and had a different disruption pattern on the muscle fibres under scanning electron micrograph. These results suggest that the cold-adapted collagenolytic protease MCP-01 may be promising for use as a meat tenderizer at low and moderate temperatures. PMID:23442615

  7. Development of an Agrobacterium-mediated transformation system for the cold-adapted fungi Pseudogymnoascus destructans and P. pannorum.

    PubMed

    Zhang, Tao; Ren, Ping; Chaturvedi, Vishnu; Chaturvedi, Sudha

    2015-08-01

    The mechanisms of cold adaptation by fungi remain unknown. This topic is of high interest due to the emergence of white-nose syndrome (WNS), a skin infection of hibernating bats caused by Pseudogymnoascus destructans (Pd). Recent studies indicated that apart from Pd, there is an abundance of other Pseudogymnoascus species in the hibernacula soil. We developed an Agrobacterium tumefaciens-mediated transformation (ATMT) system for Pd and a related fungus Pseudogymnoascus pannorum (Pp) to advance experimental studies. URE1 gene encoding the enzyme urease was used as an easy to screen marker to facilitate molecular genetic analyses. A Uracil-Specific Excision Reagent (USER) Friendly pRF-HU2 vector containing Pd or Pp ure1::hygromycin (HYG) disruption cassette was introduced into A. tumefaciens AGL-1 cells by electroporation and the resulting strains were co-cultivated with conidia of Pd or Pp for various durations and temperatures to optimize the ATMT system. Overall, 680 Pd (0.006%) and 1800 Pp (0.018%) transformants were obtained from plating of 10(7) conidia; their recoveries were strongly correlated with the length of the incubation period (96h for Pd; 72h for Pp) and with temperature (15-18°C for Pd; 25°C for Pp). The homologous recombination in transformants was 3.1% for Pd and 16.7% for Pp. The availability of a standardized ATMT system would allow future molecular genetic analyses of Pd and related cold-adapted fungi. PMID:26051491

  8. Development of an Agrobacterium-mediated transformation system for the cold-adapted fungi Pseudogymnoascus destructans and P. pannorum.

    PubMed

    Zhang, Tao; Ren, Ping; Chaturvedi, Vishnu; Chaturvedi, Sudha

    2015-08-01

    The mechanisms of cold adaptation by fungi remain unknown. This topic is of high interest due to the emergence of white-nose syndrome (WNS), a skin infection of hibernating bats caused by Pseudogymnoascus destructans (Pd). Recent studies indicated that apart from Pd, there is an abundance of other Pseudogymnoascus species in the hibernacula soil. We developed an Agrobacterium tumefaciens-mediated transformation (ATMT) system for Pd and a related fungus Pseudogymnoascus pannorum (Pp) to advance experimental studies. URE1 gene encoding the enzyme urease was used as an easy to screen marker to facilitate molecular genetic analyses. A Uracil-Specific Excision Reagent (USER) Friendly pRF-HU2 vector containing Pd or Pp ure1::hygromycin (HYG) disruption cassette was introduced into A. tumefaciens AGL-1 cells by electroporation and the resulting strains were co-cultivated with conidia of Pd or Pp for various durations and temperatures to optimize the ATMT system. Overall, 680 Pd (0.006%) and 1800 Pp (0.018%) transformants were obtained from plating of 10(7) conidia; their recoveries were strongly correlated with the length of the incubation period (96h for Pd; 72h for Pp) and with temperature (15-18°C for Pd; 25°C for Pp). The homologous recombination in transformants was 3.1% for Pd and 16.7% for Pp. The availability of a standardized ATMT system would allow future molecular genetic analyses of Pd and related cold-adapted fungi.

  9. Tenderization effect of cold-adapted collagenolytic protease MCP-01 on beef meat at low temperature and its mechanism.

    PubMed

    Zhao, Guo-Yan; Zhou, Ming-Yang; Zhao, Hui-Lin; Chen, Xiu-Lan; Xie, Bin-Bin; Zhang, Xi-Ying; He, Hai-Lun; Zhou, Bai-Cheng; Zhang, Yu-Zhong

    2012-10-15

    The enzymes currently used to increase meat tenderness are all mesophilic or thermophilic proteases. This study provides insight into the tenderization effect and the mechanism of a cold-adapted collagenolytic enzyme MCP-01 on beef meat at low temperatures. MCP-01 (10 U of caseinolytic activity) reduced the meat shear force by 23% and increased the relative myofibrillar fragmentation index of the meat by 91.7% at 4 °C, and it also kept the fresh colour and moisture of the meat. Compared to the commercially used tenderizers papain and bromelain, MCP-01 showed a unique tenderization mechanism. MCP-01 had a strong selectivity for degrading collagen at 4 °C, showed a distinct digestion pattern on the myofibrillar proteins, and had a different disruption pattern on the muscle fibres under scanning electron micrograph. These results suggest that the cold-adapted collagenolytic protease MCP-01 may be promising for use as a meat tenderizer at low and moderate temperatures.

  10. The ability of a cold-adapted Rhodotorula mucilaginosa strain from Tibet to control blue mold in pear fruit.

    PubMed

    Hu, Hao; Yan, Fujie; Wilson, Charles; Shen, Qing; Zheng, Xiaodong

    2015-12-01

    Cold-adapted yeasts were isolated from soil samples collected in Tibet and evaluated as potential biocontrol agents against blue mold (Penicillium expansum) of pear fruit in cold storage. YC1, an isolate identified as Rhodotorula mucilaginosa, was found to exhibit the greatest biocontrol activity among the different isolates that were screened. A washed cell suspension of YC1 exhibited the best biocontrol activity among three different preparations that were used in the current study. A concentration of 10(8) cells/ml reduced the incidence of decay to 35 %, compared to the control where decay incidence was 100 %. A higher intracellular level of trehalose and a higher proportion of polyunsaturated acids present in YC1, was associated with increased the tolerance of this strain to low temperatures, relative to the other strains that were evaluated. The increased tolerance to low temperature allowed the YC1 strain of yeast to more effectively compete for nutrients and space in wounded pear fruit that had been inoculated with spores of P. expansum and placed in cold storage. The present study demonstrated the ability to select cold-adapted yeasts from cold climates and use them as biocontrol agents of postharvest diseases of fruit placed in cold storage. PMID:26454432

  11. Production of cold-adapted amylase by marine bacterium Wangia sp. C52: optimization, modeling, and partial characterization.

    PubMed

    Liu, Jianguo; Zhang, Zhiqiang; Liu, Zhiqiang; Zhu, Hu; Dang, Hongyue; Lu, Jianren; Cui, Zhanfeng

    2011-10-01

    The aim of this work was to optimize the fermentation parameters in the shake-flask culture of marine bacterium Wangia sp. C52 to increase cold-adapted amylase production using two statistical experimental methods including Plackett-Burman design, which was applied to find the key ingredients for the best medium composition, and response surface methodology, which was used to determine the optimal concentrations of these components. The results showed starch, tryptone, and initial pH had significant effects on the cold-adapted amylase production. A central composite design was then employed to further optimize these three factors. The experimental results indicated that the optimized composition of medium was 6.38 g  L(-1) starch, 33.84 g  L(-1) tryptone, 3.00 g  L(-1) yeast extract, 30 g  L(-1) NaCl, 0.60 g  L(-1) MgSO(4) and 0.56 g  L(-1) CaCl(2). The optimized cultivation conditions for amylase production were pH 7.18, a temperature of 20°C, and a shaking speed of 180 rpm. Under the proposed optimized conditions, the amylase experimental yield (676.63 U  mL(-1)) closely matched the yield (685.60 U  mL(-1)) predicted by the statistical model. The optimization of the medium contributed to tenfold higher amylase production than that of the control in shake-flask experiments.

  12. Structural flexibility of the heme cavity in the cold-adapted truncated hemoglobin from the Antarctic marine bacterium Pseudoalteromonas haloplanktis TAC125.

    PubMed

    Giordano, Daniela; Pesce, Alessandra; Boechi, Leonardo; Bustamante, Juan Pablo; Caldelli, Elena; Howes, Barry D; Riccio, Alessia; di Prisco, Guido; Nardini, Marco; Estrin, Dario; Smulevich, Giulietta; Bolognesi, Martino; Verde, Cinzia

    2015-08-01

    Truncated hemoglobins build one of the three branches of the globin protein superfamily. They display a characteristic two-on-two α-helical sandwich fold and are clustered into three groups (I, II and III) based on distinct structural features. Truncated hemoglobins are present in eubacteria, cyanobacteria, protozoa and plants. Here we present a structural, spectroscopic and molecular dynamics characterization of a group-II truncated hemoglobin, encoded by the PSHAa0030 gene from Pseudoalteromonas haloplanktis TAC125 (Ph-2/2HbO), a cold-adapted Antarctic marine bacterium hosting one flavohemoglobin and three distinct truncated hemoglobins. The Ph-2/2HbO aquo-met crystal structure (at 2.21 Å resolution) shows typical features of group-II truncated hemoglobins, namely the two-on-two α-helical sandwich fold, a helix Φ preceding the proximal helix F, and a heme distal-site hydrogen-bonded network that includes water molecules and several distal-site residues, including His(58)CD1. Analysis of Ph-2/2HbO by electron paramagnetic resonance, resonance Raman and electronic absorption spectra, under varied solution conditions, shows that Ph-2/2HbO can access diverse heme ligation states. Among these, detection of a low-spin heme hexa-coordinated species suggests that residue Tyr(42)B10 can undergo large conformational changes in order to act as the sixth heme-Fe ligand. Altogether, the results show that Ph-2/2HbO maintains the general structural features of group-II truncated hemoglobins but displays enhanced conformational flexibility in the proximity of the heme cavity, a property probably related to the functional challenges, such as low temperature, high O2 concentration and low kinetic energy of molecules, experienced by organisms living in the Antarctic environment.

  13. Crystallization and preliminary X-ray diffraction analysis of a cold-adapted catalase from Vibrio salmonicida

    SciTech Connect

    Riise, Ellen Kristin; Lorentzen, Marit Sjo; Helland, Ronny; Willassen, Nils Peder

    2006-01-01

    Monoclinic (P2{sub 1}) crystals of a His-tagged form of V. salmonicida catalase without cofactor diffract X-rays to 1.96 Å. Catalase (EC 1.11.1.6) catalyses the breakdown of hydrogen peroxide to water and molecular oxygen. Recombinant Vibrio salmonicida catalase (VSC) possesses typical cold-adapted features, with higher catalytic efficiency, lower thermal stability and a lower temperature optimum than its mesophilic counterpart from Proteus mirabilis. Crystals of VSC were produced by the hanging-drop vapour-diffusion method using ammonium sulfate as precipitant. The crystals belong to the monoclinic space group P2{sub 1}, with unit-cell parameters a = 98.15, b = 217.76, c = 99.28 Å, β = 110.48°. Data were collected to 1.96 Å and a molecular-replacement solution was found with eight molecules in the asymmetric unit.

  14. Purification and characterization of novel raw-starch-digesting and cold-adapted alpha-amylases from Eisenia foetida.

    PubMed

    Ueda, Mitsuhiro; Asano, Tomohiko; Nakazawa, Masami; Miyatake, Kazutaka; Inouye, Kuniyo

    2008-05-01

    Novel raw-starch-digesting and cold-adapted alpha-amylases (Amy I and Amy II) from the earthworm Eisenia foetida were purified to electrophoretically homogeneous states. The molecular weights of both purified enzymes were estimated to be 60,000 by SDS-PAGE. The enzymes were most active at pH 5.5 and 50 degrees C and stable at pH 7.0-9.0 and 50-60 degrees C. Both Amy I and II exhibited activities at 10 degrees C. The enzymes were inhibited by metal ions Cu(2+), Fe(2+), and Hg(2+), and hydrolyzed raw starch into glucose, maltose and maltotriose as end products.

  15. Cold adaptation overrides developmental regulation of sarcolipin expression in mice skeletal muscle: SOS for muscle-based thermogenesis?

    PubMed

    Pant, Meghna; Bal, Naresh C; Periasamy, Muthu

    2015-08-01

    Neonatal mice have a greater thermogenic need than adult mice and may require additional means of heat production, other than the established mechanism of brown adipose tissue (BAT). We and others recently discovered a novel mediator of skeletal muscle-based thermogenesis called sarcolipin (SLN) that acts by uncoupling sarcoendoplasmic reticulum Ca(2+)-ATPase (SERCA). In addition, we have shown that SLN expression is downregulated during neonatal development in rats. In this study we probed two questions: (1) is SLN expression developmentally regulated in neonatal mice?; and (2) if so, will cold adaptation override this? Our data show that SLN expression is higher during early neonatal stages and is gradually downregulated in fast twitch skeletal muscles. Interestingly, we demonstrate that cold acclimation of neonatal mice can prevent downregulation of SLN expression. This observation suggests that SLN-mediated thermogenesis can be recruited to a greater extent during extreme physiological need, in addition to BAT.

  16. Identification and characterization of a novel cold-adapted esterase from a metagenomic library of mountain soil.

    PubMed

    Ko, Kyong-Cheol; Rim, Soon-Ok; Han, Yunjon; Shin, Bong Seok; Kim, Geun-Joong; Choi, Jong Hyun; Song, Jae Jun

    2012-05-01

    A novel lipolytic enzyme was isolated from a metagenomic library after demonstration of lipolytic activity on an LB agar plate containing 1% (w/v) tributyrin. A novel esterase gene (estIM1), encoding a lipolytic enzyme (EstIM1), was cloned using a shotgun method from a pFosEstIM1 clone of the metagenomic library, and the enzyme was characterized. The estIM1 gene had an open reading frame (ORF) of 936 base pairs and encoded a protein of 311 amino acids with a molecular mass 34 kDa and a pI value of 4.32. The deduced amino acid sequence was 62% identical to that of an esterase from an uncultured bacterium (ABQ11271). The amino acid sequence indicated that EstIM1 was a member of the family IV of lipolytic enzymes, all of which contain a GDSAG motif shared with similar enzymes of lactic acid microorganisms. EstIM1 was active over a temperature range of 1-50°C, at alkaline pH. The activation energy for hydrolysis of p-nitrophenyl propionate was 1.04 kcal/mol, within a temperature range of 1-40°C. The activity of EstIM1 was about 60% of maximal even at 1°C, suggesting that EstIM1 is efficiently cold-adapted. Further characterization of this cold-adapted enzyme indicated that the esterase may be very valuable in industrial applications.

  17. Glycine-rich RNA-binding proteins are functionally conserved in Arabidopsis thaliana and Oryza sativa during cold adaptation process

    PubMed Central

    Kim, Joo Yeol; Kim, Won Yong; Kwak, Kyung Jin; Oh, Seung Han; Han, Yeon Soo; Kang, Hunseung

    2010-01-01

    Contrary to the increasing amount of knowledge regarding the functional roles of glycine-rich RNA-binding proteins (GRPs) in Arabidopsis thaliana in stress responses, the physiological functions of GRPs in rice (Oryza sativa) currently remain largely unknown. In this study, the functional roles of six OsGRPs from rice on the growth of E. coli and plants under cold or freezing stress conditions have been evaluated. Among the six OsGRPs investigated, OsGRP1, OsGRP4, and OsGRP6 were shown to have the ability to complement cold-sensitive BX04 E. coli mutant cells under low temperature conditions, and this complementation ability was correlated closely with their DNA- and RNA-melting abilities. Moreover, OsGRP1 and OsGRP4 rescued the growth-defect of a cold-sensitive Arabidopsis grp7 mutant plant under cold and freezing stress, and OsGRP6 conferred freezing tolerance in the grp7 mutant plant, in which the expression of AtGRP7 was suppressed and is sensitive to cold and freezing stresses. OsGRP4 and OsGRP6 complemented the defect in mRNA export from the nucleus to the cytoplasm in grp7 mutants during cold stress. Considering that AtGRP7 confers freezing tolerance in plants and harbours RNA chaperone activity during the cold adaptation process, the results of the present study provide evidence that GRPs in rice and Arabidopsis are functionally conserved, and also suggest that GRPs perform a function as RNA chaperones during the cold adaptation process in monocotyledonous plants, as well as in dicotyledonous plants. PMID:20231330

  18. Exploring the Antarctic soil metagenome as a source of novel cold-adapted enzymes and genetic mobile elements.

    PubMed

    Berlemont, Renaud; Pipers, Delphine; Delsaute, Maud; Angiono, Federico; Feller, Georges; Galleni, Moreno; Power, Pablo

    2011-01-01

    Metagenomic library PP1 was obtained from Antarctic soil samples. Both functional and genotypic metagenomic screening were used for the isolation of novel cold-adapted enzymes with potential applications, and for the detection of genetic elements associated with gene mobilization, respectively. Fourteen lipase/esterase-, 14 amylase-, 3 protease-, and 11 cellulase-producing clones were detected by activity-driven screening, with apparent maximum activities around 35 °C for both amylolytic and lipolytic enzymes, and 35-55 °C for cellulases, as observed for other cold-adapted enzymes. However, the behavior of at least one of the studied cellulases is more compatible to that observed for mesophilic enzymes. These enzymes are usually still active at temperatures above 60 °C, probably resulting in a psychrotolerant behavior in Antarctic soils. Metagenomics allows to access novel genes encoding for enzymatic and biophysic properties from almost every environment with potential benefits for biotechnological and industrial applications. Only intI- and tnp-like genes were detected by PCR, encoding for proteins with 58-86 %, and 58-73 % amino acid identity with known entries, respectively. Two clones, BAC 27A-9 and BAC 14A-5, seem to present unique syntenic organizations, suggesting the occurrence of gene rearrangements that were probably due to evolutionary divergences within the genus or facilitated by the association with transposable elements. The evidence for genetic elements related to recruitment and mobilization of genes (transposons/integrons) in an extreme environment like Antarctica reinforces the hypothesis of the origin of some of the genes disseminated by mobile elements among "human-associated" microorganisms. PMID:21731970

  19. Predictive modeling for growth of non- and cold-adapted Listeria Monocytogenes on fresh-cut cantaloupe at different storage temperatures

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The aim of this study was to determine the growth kinetics of Listeria monocytogenes, with and without cold-adaption, on fresh-cut cantaloupe under different storage temperatures. Fresh-cut samples, spot inoculated with a four-strain cocktail of L. monocytogenes (about 3.2 log CFU/g), were exposed t...

  20. Cold Adaptation of Zinc Metalloproteases in the Thermolysin Family from Deep Sea and Arctic Sea Ice Bacteria Revealed by Catalytic and Structural Properties and Molecular Dynamics

    PubMed Central

    Xie, Bin-Bin; Bian, Fei; Chen, Xiu-Lan; He, Hai-Lun; Guo, Jun; Gao, Xiang; Zeng, Yin-Xin; Chen, Bo; Zhou, Bai-Cheng; Zhang, Yu-Zhong

    2009-01-01

    Increased conformational flexibility is the prevailing explanation for the high catalytic efficiency of cold-adapted enzymes at low temperatures. However, less is known about the structural determinants of flexibility. We reported two novel cold-adapted zinc metalloproteases in the thermolysin family, vibriolysin MCP-02 from a deep sea bacterium and vibriolysin E495 from an Arctic sea ice bacterium, and compared them with their mesophilic homolog, pseudolysin from a terrestrial bacterium. Their catalytic efficiencies, kcat/Km (10–40 °C), followed the order pseudolysin < MCP-02 < E495 with a ratio of ∼1:2:4. MCP-02 and E495 have the same optimal temperature (Topt, 57 °C, 5 °C lower than pseudolysin) and apparent melting temperature (Tm = 64 °C, ∼10 °C lower than pseudolysin). Structural analysis showed that the slightly lower stabilities resulted from a decrease in the number of salt bridges. Fluorescence quenching experiments and molecular dynamics simulations showed that the flexibilities of the proteins were pseudolysin < MCP-02 < E495, suggesting that optimization of flexibility is a strategy for cold adaptation. Molecular dynamics results showed that the ordinal increase in flexibility from pseudolysin to MCP-02 and E495, especially the increase from MCP-02 to E495, mainly resulted from the decrease of hydrogen-bond stability in the dynamic structure, which was due to the increase in asparagine, serine, and threonine residues. Finally, a model for the cold adaptation of MCP-02 and E495 was proposed. This is the first report of the optimization of hydrogen-bonding dynamics as a strategy for cold adaptation and provides new insights into the structural basis underlying conformational flexibility. PMID:19181663

  1. Structural studies of a cold-adapted dimeric β-D-galactosidase from Paracoccus sp. 32d.

    PubMed

    Rutkiewicz-Krotewicz, Maria; Pietrzyk-Brzezinska, Agnieszka J; Sekula, Bartosz; Cieśliński, Hubert; Wierzbicka-Woś, Anna; Kur, Józef; Bujacz, Anna

    2016-09-01

    The crystal structure of a novel dimeric β-D-galactosidase from Paracoccus sp. 32d (ParβDG) was solved in space group P212121 at a resolution of 2.4 Å by molecular replacement with multiple models using the BALBES software. This enzyme belongs to glycoside hydrolase family 2 (GH2), similar to the tetrameric and hexameric β-D-galactosidases from Escherichia coli and Arthrobacter sp. C2-2, respectively. It is the second known structure of a cold-active GH2 β-galactosidase, and the first in the form of a functional dimer, which is also present in the asymmetric unit. Cold-adapted β-D-galactosidases have been the focus of extensive research owing to their utility in a variety of industrial technologies. One of their most appealing applications is in the hydrolysis of lactose, which not only results in the production of lactose-free dairy, but also eliminates the `sandy effect' and increases the sweetness of the product, thus enhancing its quality. The determined crystal structure represents the five-domain architecture of the enzyme, with its active site located in close vicinity to the dimer interface. To identify the amino-acid residues involved in the catalytic reaction and to obtain a better understanding of the mechanism of action of this atypical β-D-galactosidase, the crystal structure in complex with galactose (ParβDG-Gal) was also determined. The catalytic site of the enzyme is created by amino-acid residues from the central domain 3 and from domain 4 of an adjacent monomer. The crystal structure of this dimeric β-D-galactosidase reveals significant differences in comparison to other β-galactosidases. The largest difference is in the fifth domain, named Bgal_windup domain 5 in ParβDG, which contributes to stabilization of the functional dimer. The location of this domain 5, which is unique in size and structure, may be one of the factors responsible for the creation of a functional dimer and cold-adaptation of this enzyme. PMID:27599737

  2. Structural studies of a cold-adapted dimeric β-D-galactosidase from Paracoccus sp. 32d.

    PubMed

    Rutkiewicz-Krotewicz, Maria; Pietrzyk-Brzezinska, Agnieszka J; Sekula, Bartosz; Cieśliński, Hubert; Wierzbicka-Woś, Anna; Kur, Józef; Bujacz, Anna

    2016-09-01

    The crystal structure of a novel dimeric β-D-galactosidase from Paracoccus sp. 32d (ParβDG) was solved in space group P212121 at a resolution of 2.4 Å by molecular replacement with multiple models using the BALBES software. This enzyme belongs to glycoside hydrolase family 2 (GH2), similar to the tetrameric and hexameric β-D-galactosidases from Escherichia coli and Arthrobacter sp. C2-2, respectively. It is the second known structure of a cold-active GH2 β-galactosidase, and the first in the form of a functional dimer, which is also present in the asymmetric unit. Cold-adapted β-D-galactosidases have been the focus of extensive research owing to their utility in a variety of industrial technologies. One of their most appealing applications is in the hydrolysis of lactose, which not only results in the production of lactose-free dairy, but also eliminates the `sandy effect' and increases the sweetness of the product, thus enhancing its quality. The determined crystal structure represents the five-domain architecture of the enzyme, with its active site located in close vicinity to the dimer interface. To identify the amino-acid residues involved in the catalytic reaction and to obtain a better understanding of the mechanism of action of this atypical β-D-galactosidase, the crystal structure in complex with galactose (ParβDG-Gal) was also determined. The catalytic site of the enzyme is created by amino-acid residues from the central domain 3 and from domain 4 of an adjacent monomer. The crystal structure of this dimeric β-D-galactosidase reveals significant differences in comparison to other β-galactosidases. The largest difference is in the fifth domain, named Bgal_windup domain 5 in ParβDG, which contributes to stabilization of the functional dimer. The location of this domain 5, which is unique in size and structure, may be one of the factors responsible for the creation of a functional dimer and cold-adaptation of this enzyme.

  3. Cloning, expression, purification, and characterization of cold-adapted α-amylase from Pseudoalteromonas arctica GS230.

    PubMed

    Lu, Mingsheng; Wang, Shujun; Fang, Yaowei; Li, Huangzhong; Liu, Shu; Liu, Hongfei

    2010-11-01

    A cold-adapted α-amylase (ParAmy) gene from Pseudoalteromonas arctica GS230 was cloned, sequenced, and expressed as an N-terminus His-tag fusion protein in E. coli. A recombinant protein was produced and purified with DEAE-sepherose ion exchange chromatography and Ni affinity chromatography. The molecular weight of ParAmy was estimated to be 55 KDa with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). With an optimum temperature for activity 30 °C, ParAmy showed 34.5% of maximum activity at 0 °C and its activity decreased sharply at above 40 °C. ParAmy was stable in the range of pH 7-8.5 at 30 °C for 1 h. ParAmy was activated by Mn(2+), K(+) and Na(+), and inhibited by Hg(2+), Cu(2+), and Fe(3+). N-Bromosuccinimid showed a significant repressive effect on enzyme activity. The K (m) and V (max) values of the α-amylase for soluble starch were 7.28 mg/mL and 13.07 mg/mL min, respectively. This research suggests that Paramy has a good potential to be a cold-stable and alkalitolerant amylase in detergent industry.

  4. Serine hydroxymethyltransferase from the cold adapted microorganism Psychromonas ingrahamii: a low temperature active enzyme with broad substrate specificity.

    PubMed

    Angelaccio, Sebastiana; Florio, Rita; Consalvi, Valerio; Festa, Guido; Pascarella, Stefano

    2012-01-01

    Serine hydroxymethyltransferase from the psychrophilic microorganism Psychromonas ingrahamii was expressed in Escherichia coli and purified as a His-tag fusion protein. The enzyme was characterized with respect to its spectroscopic, catalytic, and thermodynamic properties. The properties of the psychrophilic enzyme have been contrasted with the characteristics of the homologous counterpart from E. coli, which has been structurally and functionally characterized in depth and with which it shares 75% sequence identity. Spectroscopic measures confirmed that the psychrophilic enzyme displays structural properties almost identical to those of the mesophilic counterpart. At variance, the P. ingrahamii enzyme showed decreased thermostability and high specific activity at low temperature, both of which are typical features of cold adapted enzymes. Furthermore, it was a more efficient biocatalyst compared to E. coli serine hydroxymethyltransferase (SHMT) particularly for side reactions. Many β-hydroxy-α-amino acids are SHMT substrates and represent important compounds in the synthesis of pharmaceuticals, agrochemicals and food additives. Thanks to these attractive properties, this enzyme could have a significant potential for biotechnological applications.

  5. Characterization and biodegradation kinetics of a new cold-adapted carbamazepine-degrading bacterium, Pseudomonas sp. CBZ-4.

    PubMed

    Li, Ang; Cai, Rui; Di, Cui; Qiu, Tian; Pang, Changlong; Yang, Jixian; Ma, Fang; Ren, Nanqi

    2013-11-01

    Carbamazepine is frequently detected in waters and hardly eliminated during conventional wastewater treatment processes due to its complicated chemical structure and resistance to biodegradation. A carbamazepine-degrading bacterium named CBZ-4 was isolated at a low temperature (10 degreeC) from activated sludge in a municipal wastewater treatment plant. Strain CBZ-4, which can use carbamazepine as its sole source of carbon and energy, was identified as Pseudomonas sp. by the 16S rRNA gene sequence. The composition and percentage of fatty acids, which can reveal the cold-adaptation mechanism of strain CBZ-4, were determined. Strain CBZ-4 can effectively degrade carbamazepine at optimal conditions: pH 7.0, 10 degreeC, 150 r/min rotation speed, and 13% inoculation volume. The average removal rate of carbamazepine was 46.6% after 144 hr of incubation. The biodegradation kinetics of carbamazepine by CBZ-4 was fitted via the Monod model. Vmax and Ks were found to be 0.0094 hr-1 and 32.5 mg/L, respectively. PMID:24552057

  6. Is there metabolic cold adaptation in terrestrial ectotherms? Exploring latitudinal compensation in the invasive snail Cornu aspersum.

    PubMed

    Gaitán-Espitia, Juan Diego; Nespolo, Roberto

    2014-07-01

    Lower temperatures, extreme seasonality and shorter growing seasons at higher latitudes are expected to cause a decline in metabolic rates and annual growth rates of ectotherms. If a reduction in the rates of these biological processes involves a reduction in fitness, then organisms may evolve compensatory responses for the constraints imposed by high-latitude habitats. To test the existence of a latitudinal compensation in ectotherms, we used a common-garden experiment to investigate the extent to which the level of energy turnover (measured as standard metabolic rate, SMR) and the energy budget (energy allocation to growth) are affected by climatic constraints in three populations of the land snail Cornu aspersum, distributed across a latitudinal gradient of 1300 km in Chile. Our results did not support the existence of a latitudinal compensation in metabolic rates (metabolic cold adaptation). However, there was a countergradient variation (CnGV) for growth rate in which the highest latitudinal population exhibited greater growth rates than their counterparts from lower latitudes. Surprisingly, this CnGV pattern was accompanied by a lower apparent dry-matter digestibility, which could highlight a differential assimilation of ingested nutrients into somatic tissue, revealing enhanced growth efficiency in snails from the highest latitudinal habitat. Our evidence highlights that adjustments in energy allocation to the digestive machinery and to protein storage could act as a latitudinal compensation for enhanced growth efficiency in snails from the highest latitudinal population.

  7. Protective responses in mice to vaccination with multiply administered cold-adapted influenza vaccine reassortants and wild-type viruses.

    PubMed

    Romanova, J R; Tannock, G A; Alexandrova, G I

    1997-01-01

    Protective responses to influenza vaccine reassortants derived from the cold-adapted (ca) donor strains A/Leningrad/134/17/57 and B/USSR/60/69 and wild-type epidemic viruses were studied in two strains of mice. Preliminary experiments revealed that, when mixtures of three viruses were inoculated intranasally to mice with 50 microliters containing 10(6) EID50 per 200 microliters (10(5.4) EID50 per mouse), interference between strains did not occur. However, interference with the growth of the influenza reassortant B/60/32/R took place if its concentration in the mixture was reduced to 10(5) (10(4.4) per mouse) or if it was inoculated at 10(6) EID50 (10(5.4) per mouse) in the presence of the influenza reassortant R/34 and two other influenza A epidemic strains; interference was unrelated to serological responses to infection with B/60/32/R. Despite evidence of interference, mice inoculated with the same mixtures in two identical doses, three weeks apart, were able to clear a challenge from each of seven homotypic and heterotypic influenza A and B strains. Heterotypic clearance of influenza A challenge viruses was greater following mixed infection, indicating that common determinants within the surface antigen glycoproteins contributed to immune responses which were broader than could be expected to be induced by parenteral vaccination.

  8. A novel cold-adapted and glucose-tolerant GH1 β-glucosidase from Exiguobacterium antarcticum B7.

    PubMed

    Crespim, Elaine; Zanphorlin, Letícia M; de Souza, Flavio H M; Diogo, José A; Gazolla, Alex C; Machado, Carla B; Figueiredo, Fernanda; Sousa, Amanda S; Nóbrega, Felipe; Pellizari, Vivian H; Murakami, Mário T; Ruller, Roberto

    2016-01-01

    A novel GH1 β-glucosidase (EaBgl1A) from a bacterium isolated from Antarctica soil samples was recombinantly overexpressed in Escherichia coli cells and characterized. The enzyme showed unusual pH dependence with maximum activity at neutral pH and retention of high catalytic activity in the pH range 6 to 9, indicating a catalytic machinery compatible with alkaline conditions. EaBgl1A is also a cold-adapted enzyme, exhibiting activity in the temperature range from 10 to 40°C with optimal activity at 30°C, which allows its application in industrial processes using low temperatures. Kinetic characterization revealed an enzymatic turnover (Kcat) of 6.92s(-1) (cellobiose) and 32.98s(-1) (pNPG) and a high tolerance for product inhibition, which is an extremely desirable feature for biotechnological purposes. Interestingly, the enzyme was stimulated by up to 200 mM glucose, whereas the commercial cocktails tested were found fully inhibited at this concentration. These properties indicate EaBgl1A as a promising biocatalyst for biotechnological applications where low temperatures are required.

  9. An exceptionally cold-adapted alpha-amylase from a metagenomic library of a cold and alkaline environment.

    PubMed

    Vester, Jan Kjølhede; Glaring, Mikkel Andreas; Stougaard, Peter

    2015-01-01

    A cold-active α-amylase, AmyI3C6, identified by a functional metagenomics approach was expressed in Escherichia coli and purified to homogeneity. Sequence analysis showed that the AmyI3C6 amylase was similar to α-amylases from the class Clostridia and revealed classical characteristics of cold-adapted enzymes, as did comparison of the kinetic parameters K m and k cat to a mesophilic α-amylase. AmyI3C6 was shown to be heat-labile. Temperature optimum was at 10-15 °C, and more than 70 % of the relative activity was retained at 1 °C. The pH optimum of AmyI3C6 was at pH 8-9, and the enzyme displayed activity in two commercial detergents tested, suggesting that the AmyI3C6 α-amylase may be useful as a detergent enzyme in environmentally friendly, low-temperature laundry processes.

  10. Purification, characterization, and gene cloning of a cold-adapted thermolysin-like protease from Halobacillus sp. SCSIO 20089.

    PubMed

    Yang, Jian; Li, Jie; Mai, Zhimao; Tian, Xinpeng; Zhang, Si

    2013-06-01

    Marine sediment is a distinctive habitat of cold enzyme producing bacteria. A protease producing strain Halobacillus sp. SCSIO 20089 was isolated from a marine sediment of South China Sea. Using chromatographic techniques, the extracellular protease was purified to homogeneity from the culture supernatant. The purified protease exhibited maximal activity at 30°C, pH 8.0, and remained more than 20% of its activity at 0°C. Its activation energy was calculated to be 34.4 kJ/mol, suggesting it is a cold-adapted protease. Based on the N-terminal amino acid sequence of the purified enzyme, full gene encoding the enzyme was obtained by combination of degenerate primer PCR and hiTAIL-PCR. The deduced amino acid sequence showed 57% and 52% identity with mesothermal and thermophilic protease in thermolysin family respectively. All these indicate the enzyme is a unique cold-active thermolysin-like protease with potential in both basic research and industrial application areas.

  11. Distribution of cold adaptation proteins in microbial mats in Lake Joyce, Antarctica: Analysis of metagenomic data by using two bioinformatics tools.

    PubMed

    Koo, Hyunmin; Hakim, Joseph A; Fisher, Phillip R E; Grueneberg, Alexander; Andersen, Dale T; Bej, Asim K

    2016-01-01

    In this study, we report the distribution and abundance of cold-adaptation proteins in microbial mat communities in the perennially ice-covered Lake Joyce, located in the McMurdo Dry Valleys, Antarctica. We have used MG-RAST and R code bioinformatics tools on Illumina HiSeq2000 shotgun metagenomic data and compared the filtering efficacy of these two methods on cold-adaptation proteins. Overall, the abundance of cold-shock DEAD-box protein A (CSDA), antifreeze proteins (AFPs), fatty acid desaturase (FAD), trehalose synthase (TS), and cold-shock family of proteins (CSPs) were present in all mat samples at high, moderate, or low levels, whereas the ice nucleation protein (INP) was present only in the ice and bulbous mat samples at insignificant levels. Considering the near homogeneous temperature profile of Lake Joyce (0.08-0.29 °C), the distribution and abundance of these proteins across various mat samples predictively correlated with known functional attributes necessary for microbial communities to thrive in this ecosystem. The comparison of the MG-RAST and the R code methods showed dissimilar occurrences of the cold-adaptation protein sequences, though with insignificant ANOSIM (R = 0.357; p-value = 0.012), ADONIS (R(2) = 0.274; p-value = 0.03) and STAMP (p-values = 0.521-0.984) statistical analyses. Furthermore, filtering targeted sequences using the R code accounted for taxonomic groups by avoiding sequence redundancies, whereas the MG-RAST provided total counts resulting in a higher sequence output. The results from this study revealed for the first time the distribution of cold-adaptation proteins in six different types of microbial mats in Lake Joyce, while suggesting a simpler and more manageable user-defined method of R code, as compared to a web-based MG-RAST pipeline. PMID:26578243

  12. Distribution of cold adaptation proteins in microbial mats in Lake Joyce, Antarctica: Analysis of metagenomic data by using two bioinformatics tools.

    PubMed

    Koo, Hyunmin; Hakim, Joseph A; Fisher, Phillip R E; Grueneberg, Alexander; Andersen, Dale T; Bej, Asim K

    2016-01-01

    In this study, we report the distribution and abundance of cold-adaptation proteins in microbial mat communities in the perennially ice-covered Lake Joyce, located in the McMurdo Dry Valleys, Antarctica. We have used MG-RAST and R code bioinformatics tools on Illumina HiSeq2000 shotgun metagenomic data and compared the filtering efficacy of these two methods on cold-adaptation proteins. Overall, the abundance of cold-shock DEAD-box protein A (CSDA), antifreeze proteins (AFPs), fatty acid desaturase (FAD), trehalose synthase (TS), and cold-shock family of proteins (CSPs) were present in all mat samples at high, moderate, or low levels, whereas the ice nucleation protein (INP) was present only in the ice and bulbous mat samples at insignificant levels. Considering the near homogeneous temperature profile of Lake Joyce (0.08-0.29 °C), the distribution and abundance of these proteins across various mat samples predictively correlated with known functional attributes necessary for microbial communities to thrive in this ecosystem. The comparison of the MG-RAST and the R code methods showed dissimilar occurrences of the cold-adaptation protein sequences, though with insignificant ANOSIM (R = 0.357; p-value = 0.012), ADONIS (R(2) = 0.274; p-value = 0.03) and STAMP (p-values = 0.521-0.984) statistical analyses. Furthermore, filtering targeted sequences using the R code accounted for taxonomic groups by avoiding sequence redundancies, whereas the MG-RAST provided total counts resulting in a higher sequence output. The results from this study revealed for the first time the distribution of cold-adaptation proteins in six different types of microbial mats in Lake Joyce, while suggesting a simpler and more manageable user-defined method of R code, as compared to a web-based MG-RAST pipeline.

  13. Purification and characterization of a novel cold-adapted phytase from Rhodotorula mucilaginosa strain JMUY14 isolated from Antarctic.

    PubMed

    Yu, Peng; Wang, Xue-Ting; Liu, Jing-Wen

    2015-08-01

    A yeast producing a cold-adapted phytase was isolated from Antarctic deep-sea sediment and identified as a Rhodotorula mucilaginosa strain JMUY14 of basidiomycetous yeasts. It was cultured in fermentation optimized by a response surface methodology based on the Box-Behnken design. The maximum activity of phytase reached 205.447 U ml(-1), which was close to the predicted value of 201.948 U ml(-1) and approximately 3.4 times higher than its initial activity. The extracellular phytase was purified by 15.2-fold to homogeneity with a specific activity of 31,635 U mg(-1) by (NH4 )2 SO4 precipitation, and a combination of DEAE Sepharose Fast Flow, SP Sepharose Fast Flow, and Sephadex G-100. The molecular weight of the purified enzyme was estimated to be 63 kDa and its pI was 4.33. Its optimal temperature and pH were 50 °C and 5.0, respectively. Its activity was 85% at 37 °C, and showed good stability at pH 3.0 ∼ 7.0. When compared with mesophilic counterparts, the phytase not only exhibited a higher activity during 20 ∼ 30 °C but also had a low Km (247 µM) and high kcat (1394 s(-1)). The phytase activity was slightly stimulated in the presence of Mg(2+), Fe(2+), Fe(3+), K(+), Na(+), Ca(2+), EDTA, and EGTA and moderately inhibited by Cu(2+), Zn(2+), Mn(2+), Ag(+), PMSF, SDS, and phenylgloxal hydrate. It was resistant to both pepsin and trypsin. Since the phytase produced by the R. mucilaginosa JMUY14 showed a high specific activity, good pH stability, strong protease resistance, and high activity at low temperature, it has great potential for feed applications, especially in aquaculture. PMID:25727311

  14. A single mutation Gln142Lys doubles the catalytic activity of VPR, a cold adapted subtilisin-like serine proteinase.

    PubMed

    Óskarsson, Kristinn R; Nygaard, Mads; Ellertsson, Brynjar Ö; Thorbjarnardottir, Sigríður H; Papaleo, Elena; Kristjánsson, Magnús M

    2016-10-01

    Structural comparisons of the cold adapted subtilase VPR and its thermophilic homologue, aqualysin I (AQUI) indicated the presence of additional salt bridges in the latter. Few of those appear to contribute significantly to thermal stability of AQUI. This includes a putative salt bridge between residues Lys142 and Glu172 as its deletion did not have any significant effect on its stability or activity (Jónsdóttir et al. (2014)). Insertion of this putative salt bridge into the structure of VPR, in a double mutant (VPRΔC_Q142K/S172E), however was detrimental to the stability of the enzyme. Incorporation of either the Q142K or S172E mutations into VPR, were found to significantly affect the catalytic properties of the enzyme. The single mutation Q142K was highly effective, as it increased the kcat and kcat/Km more than twofold. When the Q142K mutation was inserted into a thermostabilized, but a low activity mutant of VPR (VPRΔC_N3P/I5P), the activity increased about tenfold in terms of kcat and kcat/Km, while retaining the stability of the mutant. Molecular dynamics simulations of the single mutants were carried out to provide structural rationale for these experimental observations. Based on root mean square fluctuation (RMSF) profiles, the two mutants were more flexible in certain regions of the structure and the Q142K mutant had the highest overall flexibility of the three enzymes. The results suggest that weakening of specific H-bonds resulting from the mutations may be propagated over some distance giving rise to higher flexibility in the active site regions of the enzyme, causing higher catalytic activity in the mutants. PMID:27456266

  15. The Genome Sequence of the psychrophilic archaeon, Methanococcoides burtonii: the Role of Genome Evolution in Cold-adaptation

    SciTech Connect

    Allen, Michelle A.; Lauro, Federico M.; Williams, Timothy J.; Burg, Dominic; Siddiqui, Khawar S.; De Francisci, David; Chong, Kevin W.Y.; Pilak, Oliver; Chew, Hwee H.; De Maere, Matthew Z.; Ting, Lily; Katrib, Marilyn; Ng, Charmaine; Sowers, Kevin R.; Galperin, Michael Y.; Anderson, Iain J.; Ivanova, Natalia; Dalin, Eileen; Martinez, Michelle; Lapidus, Alla; Hauser, Loren; Land, Miriam; Thomas, Torsten; Cavicchioli, Ricardo

    2009-04-01

    Psychrophilic archaea are abundant and perform critical roles throughout the Earth's expansive cold biosphere. Here we report the first complete genome sequence for a psychrophilic methanogenic archaeon, Methanococcoides burtonii. The genome sequence was manually annotated including the use of a five tiered Evidence Rating system that ranked annotations from Evidence Rating (ER) 1 (gene product experimentally characterized from the parent organism) to ER5 (hypothetical gene product) to provide a rapid means of assessing the certainty of gene function predictions. The genome is characterized by a higher level of aberrant sequence composition (51%) than any other archaeon. In comparison to hyper/thermophilic archaea which are subject to selection of synonymous codon usage, M. burtonii has evolved cold adaptation through a genomic capacity to accommodate highly skewed amino acid content, while retaining codon usage in common with its mesophilic Methanosarcina cousins. Polysaccharide biosynthesis genes comprise at least 3.3% of protein coding genes in the genome, and Cell wall/membrane/envelope biogenesis COG genes are over-represented. Likewise, signal transduction (COG category T) genes are over-represented and M. burtonii has a high 'IQ' (a measure of adaptive potential) compared to many methanogens. Numerous genes in these two over-represented COG categories appear to have been acquired from {var_epsilon}- and {delta}-proteobacteria, as do specific genes involved in central metabolism such as a novel B form of aconitase. Transposases also distinguish M. burtonii from other archaea, and their genomic characteristics indicate they play an important role in evolving the M. burtonii genome. Our study reveals a capacity for this model psychrophile to evolve through genome plasticity (including nucleotide skew, horizontal gene transfer and transposase activity) that enables adaptation to the cold, and to the biological and physical changes that have occurred over the

  16. The genome sequence of the psychrophilic archaeon, Methanococcoides burtonii: the role of genome evolution in cold adaptation

    SciTech Connect

    Allen, Michele A; Lauro, Federico M; Williams, Timothy J; Burg, Dominic; Siddiqui, Khawar S; DeFrancisci, Davide; Chong, Kevin WY; Pilak, Oliver; Chew, Hwee H; DeMaere, Matthew Z; Ting, Lily; Katrib, Marilyn; Ng, Charmaine; Sowers, Kevin R; Galperin, Michael Y.; Anderson, Iain; Ivanova, N; Dalin, Eileen; Martinez, Michele; Lapidus, Alla L.; Hauser, Loren John; Land, Miriam L; Thomas, Torsten; Cavicchioli, Ricardo

    2009-01-01

    Psychrophilic archaea are abundant and perform critical roles throughout the Earth's expansive cold biosphere. Here we report the first complete genome sequence for a psychrophilic methanogenic archaeon, Methanococcoides burtonii. The genome sequence was manually annotated including the use of a five-tiered evidence rating (ER) system that ranked annotations from ER1 (gene product experimentally characterized from the parent organism) to ER5 (hypothetical gene product) to provide a rapid means of assessing the certainty of gene function predictions. The genome is characterized by a higher level of aberrant sequence composition (51%) than any other archaeon. In comparison to hyper/thermophilic archaea, which are subject to selection of synonymous codon usage, M. burtonii has evolved cold adaptation through a genomic capacity to accommodate highly skewed amino-acid content, while retaining codon usage in common with its mesophilic Methanosarcina cousins. Polysaccharide biosynthesis genes comprise at least 3.3% of protein coding genes in the genome, and Cell wall, membrane, envelope biogenesis COG genes are overrepresented. Likewise, signal transduction (COG category T) genes are overrepresented and M. burtonii has a high 'IQ' (a measure of adaptive potential) compared to many methanogens. Numerous genes in these two overrepresented COG categories appear to have been acquired from - and -Proteobacteria, as do specific genes involved in central metabolism such as a novel B form of aconitase. Transposases also distinguish M. burtonii from other archaea, and their genomic characteristics indicate they have an important role in evolving the M. burtonii genome. Our study reveals a capacity for this model psychrophile to evolve through genome plasticity (including nucleotide skew, horizontal gene transfer and transposase activity) that enables adaptation to the cold, and to the biological and physical changes that have occurred over the last several thousand years as it

  17. Antarctic marine bacterium Pseudoalteromonas sp. 22b as a source of cold-adapted beta-galactosidase.

    PubMed

    Turkiewicz, Marianna; Kur, Józef; Białkowska, Aneta; Cieśliński, Hubert; Kalinowska, Halina; Bielecki, Stanisław

    2003-07-01

    The marine, psychrotolerant, rod-shaped and Gram-negative bacterium 22b (the best of 41 beta-galactosidase producers out of 107 Antarctic strains subjected to screening), classified as Pseudoalteromonas sp. based on 16S rRNA gene sequence, isolated from the alimentary tract of Antarctic krill Thyssanoessa macrura, synthesizes an intracellular cold-adapted beta-galactosidase, which efficiently hydrolyzes lactose at 0-20 degrees C, as indicated by its specific activity of 21-67 U mg(-1) of protein (11-35% of maximum activity) in this temperature range, as well as k(cat) of 157 s(-1), and k(cat)/K(m) of 47.5 mM(-1) s(-1) at 20 degrees C. The maximum enzyme synthesis (lactose as a sufficient inducer) was observed at 6 degrees C, thus below the optimum growth temperature of the bacterium (15 degrees C). The enzyme extracted from cells was purified to homogeneity (25% recovery) by using the fast, three-step procedure, including affinity chromatography on PABTG-Sepharose. The enzyme is a tetramer composed of roughly 115 kDa subunits. It is maximally active at 40 degrees C (190 U mg(-1) of protein) and pH 6.0-8.0. PNPG is its preferred substrate (50% higher activity than against ONPG). The Pseudoalteromonas sp. 22b beta-galactosidase is activated by thiol compounds (70% rise in activity in the presence of 10 mM dithiotreitol), some metal ions (K(+), Na(+), Mn(2+)-40% increase, Mg(2+)-15% enhancement), and markedly inactivated by pCMB and heavy metal ions, particularly Cu(2+). Noteworthy, Ca(2+) ions do not affect the enzyme activity, and the homogeneous protein is stable at 4 degrees C for at least 30 days without any stabilizers.

  18. Oligomerization as a strategy for cold adaptation: Structure and dynamics of the GH1 β-glucosidase from Exiguobacterium antarcticum B7

    PubMed Central

    Zanphorlin, Leticia Maria; de Giuseppe, Priscila Oliveira; Honorato, Rodrigo Vargas; Tonoli, Celisa Caldana Costa; Fattori, Juliana; Crespim, Elaine; de Oliveira, Paulo Sergio Lopes; Ruller, Roberto; Murakami, Mario Tyago

    2016-01-01

    Psychrophilic enzymes evolved from a plethora of structural scaffolds via multiple molecular pathways. Elucidating their adaptive strategies is instrumental to understand how life can thrive in cold ecosystems and to tailor enzymes for biotechnological applications at low temperatures. In this work, we used X-ray crystallography, in solution studies and molecular dynamics simulations to reveal the structural basis for cold adaptation of the GH1 β-glucosidase from Exiguobacterium antarcticum B7. We discovered that the selective pressure of low temperatures favored mutations that redesigned the protein surface, reduced the number of salt bridges, exposed more hydrophobic regions to the solvent and gave rise to a tetrameric arrangement not found in mesophilic and thermophilic homologues. As a result, some solvent-exposed regions became more flexible in the cold-adapted tetramer, likely contributing to enhance enzymatic activity at cold environments. The tetramer stabilizes the native conformation of the enzyme, leading to a 10-fold higher activity compared to the disassembled monomers. According to phylogenetic analysis, diverse adaptive strategies to cold environments emerged in the GH1 family, being tetramerization an alternative, not a rule. These findings reveal a novel strategy for enzyme cold adaptation and provide a framework for the semi-rational engineering of β-glucosidases aiming at cold industrial processes. PMID:27029646

  19. Functional annotation of the mesophilic-like character of mutants in a cold-adapted enzyme by self-organising map analysis of their molecular dynamics.

    PubMed

    Fraccalvieri, Domenico; Tiberti, Matteo; Pandini, Alessandro; Bonati, Laura; Papaleo, Elena

    2012-10-01

    Multiple comparison of the Molecular Dynamics (MD) trajectories of mutants in a cold-adapted α-amylase (AHA) could be used to elucidate functional features required to restore mesophilic-like activity. Unfortunately it is challenging to identify the different dynamic behaviors and correctly relate them to functional activity by routine analysis. We here employed a previously developed and robust two-stage approach that combines Self-Organising Maps (SOMs) and hierarchical clustering to compare conformational ensembles of proteins. Moreover, we designed a novel strategy to identify the specific mutations that more efficiently convert the dynamic signature of the psychrophilic enzyme (AHA) to that of the mesophilic counterpart (PPA). The SOM trained on AHA and its variants was used to classify a PPA MD ensemble and successfully highlighted the relationships between the flexibilities of the target enzyme and of the different mutants. Moreover the local features of the mutants that mostly influence their global flexibility in a mesophilic-like direction were detected. It turns out that mutations of the cold-adapted enzyme to hydrophobic and aromatic residues are the most effective in restoring the PPA dynamic features and could guide the design of more mesophilic-like mutants. In conclusion, our strategy can efficiently extract specific dynamic signatures related to function from multiple comparisons of MD conformational ensembles. Therefore, it can be a promising tool for protein engineering.

  20. Contributions of Two-Component Regulatory Systems, Alternative σ Factors, and Negative Regulators to Listeria monocytogenes Cold Adaptation and Cold Growth

    PubMed Central

    Chan, Yvonne C.; Hu, Yuewei; Chaturongakul, Soraya; Files, Kali D.; Bowen, Barbara M.; Boor, Kathryn J.; Wiedmann, Martin

    2011-01-01

    The ability of Listeria monocytogenes to grow at refrigeration temperatures is critical for transmission of this foodborne pathogen. We evaluated the contributions of different transcriptional regulators and two-component regulatory systems to L. monocytogenes cold adaptation and cold growth. L. monocytogenes parent strain 10403S and selected isogenic null mutants in genes encoding four alternative σ factors (sigB, sigH, sigC, and sigL), two regulators of σB (rsbT and rsbV), two negative regulators (ctsR and hrcA), and 15 two-component response regulators were grown in brain heart infusion broth at 4°C with (i) a high-concentration starting inoculum (108 CFU/ml), (ii) a low-concentration starting inoculum (102 CFU/ml), and (iii) a high-concentration starting inoculum of cold-adapted cells. With a starting inoculum of 108 CFU/ml, null mutants in genes encoding selected alternative σ factors (ΔsigH, ΔsigC, and ΔsigL), a negative regulator (ΔctsR), regulators of σB (ΔrsbT and ΔrsbV), and selected two-component response regulators (ΔlisR, Δlmo1172, and Δlmo1060) had significantly reduced growth (P < 0.05) compared with the parent strain after 12 days at 4°C. The growth defect for ΔsigL was limited and was not confirmed by optical density (OD600) measurement data. With a starting inoculum of 102 CFU/ml and after monitoring growth at 4°C over 84 days, only the ΔctsR strain had a consistent but limited growth defect; the other mutant strains had either no growth defects or limited growth defects apparent at only one or two of the nine sampling points evaluated during the 84-day growth period (ΔsigB, ΔsigC, and Δlmo1172). With a 108 CFU/ml starting inoculum of cold-adapted cells, none of the mutant strains that had a growth defect when inoculation was performed with cells pregrown at 37°C had reduced growth as compared with the parent strain after 12 days at 4°C, suggesting a specific defect in the ability of these mutant strains to adapt to 4

  1. Cold shock genes cspA and cspB from Caulobacter crescentus are posttranscriptionally regulated and important for cold adaptation.

    PubMed

    Mazzon, Ricardo R; Lang, Elza A S; Silva, Carolina A P T; Marques, Marilis V

    2012-12-01

    Cold shock proteins (CSPs) are nucleic acid binding chaperones, first described as being induced to solve the problem of mRNA stabilization after temperature downshift. Caulobacter crescentus has four CSPs: CspA and CspB, which are cold induced, and CspC and CspD, which are induced only in stationary phase. In this work we have determined that the synthesis of both CspA and CspB reaches the maximum levels early in the acclimation phase. The deletion of cspA causes a decrease in growth at low temperature, whereas the strain with a deletion of cspB has a very subtle and transient cold-related growth phenotype. The cspA cspB double mutant has a slightly more severe phenotype than that of the cspA mutant, suggesting that although CspA may be more important to cold adaptation than CspB, both proteins have a role in this process. Gene expression analyses were carried out using cspA and cspB regulatory fusions to the lacZ reporter gene and showed that both genes are regulated at the transcriptional and posttranscriptional levels. Deletion mapping of the long 5'-untranslated region (5'-UTR) of each gene identified a common region important for cold induction, probably via translation enhancement. In contrast to what was reported for other bacteria, these cold shock genes have no regulatory regions downstream from ATG that are important for cold induction. This work shows that the importance of CspA and CspB to C. crescentus cold adaptation, mechanisms of regulation, and pattern of expression during the acclimation phase apparently differs in many aspects from what has been described so far for other bacteria.

  2. Biodegradation and metabolic pathway of sulfamethoxazole by Pseudomonas psychrophila HA-4, a newly isolated cold-adapted sulfamethoxazole-degrading bacterium.

    PubMed

    Jiang, Benchao; Li, Ang; Cui, Di; Cai, Rui; Ma, Fang; Wang, Yingning

    2014-05-01

    Sulfamethoxazole is a common antibiotic that is frequently detected in wastewater and surface water. This study investigated the biodegradation and metabolic pathway of sulfamethoxazole by Pseudomonas psychrophila HA-4, a cold-adapted bacterium. Strain HA-4, which uses sulfamethoxazole as its sole source of carbon and energy, was isolated at a low temperature (10 °C) and identified as P. psychrophila by physico-biochemical characterization and 16S rRNA gene sequence analysis. Strain HA-4 removed sulfamethoxazole at temperatures ranging from 5.0 °C to 30 °C, with the maximal removal rate at 10 °C. The maximal removal rate of sulfamethoxazole by strain HA-4 was 34.30 % after 192 h at 10 °C. The highest percentage of unsaturated fatty acid was determined to be 23.03 % at 10 °C, which adheres to the characteristic for cold-adapted psychrophiles and psychrotrophs. At low concentrations of sulfamethoxazole, the growth kinetics correlated well with the Haldane model. The single-substrate parameter values of sulfamethoxazole on cell growth were determined to be μ max = 0.01 h(-1), K s = 20.91 mg/l and K i = 170.60 mg/l. Additionally, the major intermediates from sulfamethoxazole biodegradation by strain HA-4, including aniline, 3-amino-5-methylisoxazole, 4-aminothiophenol and sulfanilamide, were identified by GC-MS and high-resolution mass spectrometry (HR-MS) analysis. The results demonstrate that strain HA-4 has the potential to degrade sulfamethoxazole at low temperatures. PMID:24522726

  3. Biodegradation and metabolic pathway of sulfamethoxazole by Pseudomonas psychrophila HA-4, a newly isolated cold-adapted sulfamethoxazole-degrading bacterium.

    PubMed

    Jiang, Benchao; Li, Ang; Cui, Di; Cai, Rui; Ma, Fang; Wang, Yingning

    2014-05-01

    Sulfamethoxazole is a common antibiotic that is frequently detected in wastewater and surface water. This study investigated the biodegradation and metabolic pathway of sulfamethoxazole by Pseudomonas psychrophila HA-4, a cold-adapted bacterium. Strain HA-4, which uses sulfamethoxazole as its sole source of carbon and energy, was isolated at a low temperature (10 °C) and identified as P. psychrophila by physico-biochemical characterization and 16S rRNA gene sequence analysis. Strain HA-4 removed sulfamethoxazole at temperatures ranging from 5.0 °C to 30 °C, with the maximal removal rate at 10 °C. The maximal removal rate of sulfamethoxazole by strain HA-4 was 34.30 % after 192 h at 10 °C. The highest percentage of unsaturated fatty acid was determined to be 23.03 % at 10 °C, which adheres to the characteristic for cold-adapted psychrophiles and psychrotrophs. At low concentrations of sulfamethoxazole, the growth kinetics correlated well with the Haldane model. The single-substrate parameter values of sulfamethoxazole on cell growth were determined to be μ max = 0.01 h(-1), K s = 20.91 mg/l and K i = 170.60 mg/l. Additionally, the major intermediates from sulfamethoxazole biodegradation by strain HA-4, including aniline, 3-amino-5-methylisoxazole, 4-aminothiophenol and sulfanilamide, were identified by GC-MS and high-resolution mass spectrometry (HR-MS) analysis. The results demonstrate that strain HA-4 has the potential to degrade sulfamethoxazole at low temperatures.

  4. Cold-Adapted Viral Attenuation (CAVA): Highly Temperature Sensitive Polioviruses as Novel Vaccine Strains for a Next Generation Inactivated Poliovirus Vaccine.

    PubMed

    Sanders, Barbara P; de Los Rios Oakes, Isabel; van Hoek, Vladimir; Bockstal, Viki; Kamphuis, Tobias; Uil, Taco G; Song, Yutong; Cooper, Gillian; Crawt, Laura E; Martín, Javier; Zahn, Roland; Lewis, John; Wimmer, Eckard; Custers, Jerome H H V; Schuitemaker, Hanneke; Cello, Jeronimo; Edo-Matas, Diana

    2016-03-01

    The poliovirus vaccine field is moving towards novel vaccination strategies. Withdrawal of the Oral Poliovirus Vaccine and implementation of the conventional Inactivated Poliovirus Vaccine (cIPV) is imminent. Moreover, replacement of the virulent poliovirus strains currently used for cIPV with attenuated strains is preferred. We generated Cold-Adapted Viral Attenuation (CAVA) poliovirus strains by serial passage at low temperature and subsequent genetic engineering, which contain the capsid sequences of cIPV strains combined with a set of mutations identified during cold-adaptation. These viruses displayed a highly temperature sensitive phenotype with no signs of productive infection at 37°C as visualized by electron microscopy. Furthermore, decreases in infectious titers, viral RNA, and protein levels were measured during infection at 37°C, suggesting a block in the viral replication cycle at RNA replication, protein translation, or earlier. However, at 30°C, they could be propagated to high titers (9.4-9.9 Log10TCID50/ml) on the PER.C6 cell culture platform. We identified 14 mutations in the IRES and non-structural regions, which in combination induced the temperature sensitive phenotype, also when transferred to the genomes of other wild-type and attenuated polioviruses. The temperature sensitivity translated to complete absence of neurovirulence in CD155 transgenic mice. Attenuation was also confirmed after extended in vitro passage at small scale using conditions (MOI, cell density, temperature) anticipated for vaccine production. The inability of CAVA strains to replicate at 37°C makes reversion to a neurovirulent phenotype in vivo highly unlikely, therefore, these strains can be considered safe for the manufacture of IPV. The CAVA strains were immunogenic in the Wistar rat potency model for cIPV, inducing high neutralizing antibody titers in a dose-dependent manner in response to D-antigen doses used for cIPV. In combination with the highly productive

  5. Cold-Adapted Viral Attenuation (CAVA): Highly Temperature Sensitive Polioviruses as Novel Vaccine Strains for a Next Generation Inactivated Poliovirus Vaccine

    PubMed Central

    Sanders, Barbara P.; de los Rios Oakes, Isabel; van Hoek, Vladimir; Bockstal, Viki; Kamphuis, Tobias; Uil, Taco G.; Song, Yutong; Cooper, Gillian; Crawt, Laura E.; Martín, Javier; Zahn, Roland; Lewis, John; Wimmer, Eckard; Custers, Jerome H. H. V.; Schuitemaker, Hanneke; Cello, Jeronimo; Edo-Matas, Diana

    2016-01-01

    The poliovirus vaccine field is moving towards novel vaccination strategies. Withdrawal of the Oral Poliovirus Vaccine and implementation of the conventional Inactivated Poliovirus Vaccine (cIPV) is imminent. Moreover, replacement of the virulent poliovirus strains currently used for cIPV with attenuated strains is preferred. We generated Cold-Adapted Viral Attenuation (CAVA) poliovirus strains by serial passage at low temperature and subsequent genetic engineering, which contain the capsid sequences of cIPV strains combined with a set of mutations identified during cold-adaptation. These viruses displayed a highly temperature sensitive phenotype with no signs of productive infection at 37°C as visualized by electron microscopy. Furthermore, decreases in infectious titers, viral RNA, and protein levels were measured during infection at 37°C, suggesting a block in the viral replication cycle at RNA replication, protein translation, or earlier. However, at 30°C, they could be propagated to high titers (9.4–9.9 Log10TCID50/ml) on the PER.C6 cell culture platform. We identified 14 mutations in the IRES and non-structural regions, which in combination induced the temperature sensitive phenotype, also when transferred to the genomes of other wild-type and attenuated polioviruses. The temperature sensitivity translated to complete absence of neurovirulence in CD155 transgenic mice. Attenuation was also confirmed after extended in vitro passage at small scale using conditions (MOI, cell density, temperature) anticipated for vaccine production. The inability of CAVA strains to replicate at 37°C makes reversion to a neurovirulent phenotype in vivo highly unlikely, therefore, these strains can be considered safe for the manufacture of IPV. The CAVA strains were immunogenic in the Wistar rat potency model for cIPV, inducing high neutralizing antibody titers in a dose-dependent manner in response to D-antigen doses used for cIPV. In combination with the highly productive

  6. Characterization of a pH and detergent-tolerant, cold-adapted type I pullulanase from Exiguobacterium sp. SH3.

    PubMed

    Rajaei, Sarah; Noghabi, Kambiz Akbari; Sadeghizadeh, Majid; Zahiri, Hossein Shahbani

    2015-11-01

    A pullulanase-encoding gene from psychrotrophic Exiguobacterium sp. SH3 was cloned and expressed in both E. coli and Bacillus subtilis. The functional recombinant enzyme (Pul-SH3) was purified as a His-tagged protein. Pul-SH3 was characterized to be a cold-adapted type I pullulanase with maximum activity at 45 °C. Using fluorescence spectroscopy, the melting temperature of Pul-SH3 was determined to be about 52 °C. The enzyme was able to hydrolyze pullulan, soluble starch, potato starch, and rice flour. It was exceptionally tolerant in the pH range of 4-11, exhibiting maximum activity at pH 8.5 and more than 60% of the activity in the pH range of 5-11. Being a detergent-tolerant pullulanase, Pul-SH3 retained 99, 89, and 54% of its activity at 10% concentration of Triton-X100, Tween 20, and SDS, respectively. The enzyme also exhibited an activity of 80.4 and 93.7% in the presence of two commercial detergents, Rika (7.5% v/v) and Fadisheh (2.5% w/v), respectively. The enzyme was even able to remain active by 54.5 and 85% after 10-day holding with the commercial detergents. Thermal stability of the enzyme could w on silica. Pul-SH3 with several industrially important characteristics seems desirable for cold hydrolysis of starch.

  7. Characterization of a pH and detergent-tolerant, cold-adapted type I pullulanase from Exiguobacterium sp. SH3.

    PubMed

    Rajaei, Sarah; Noghabi, Kambiz Akbari; Sadeghizadeh, Majid; Zahiri, Hossein Shahbani

    2015-11-01

    A pullulanase-encoding gene from psychrotrophic Exiguobacterium sp. SH3 was cloned and expressed in both E. coli and Bacillus subtilis. The functional recombinant enzyme (Pul-SH3) was purified as a His-tagged protein. Pul-SH3 was characterized to be a cold-adapted type I pullulanase with maximum activity at 45 °C. Using fluorescence spectroscopy, the melting temperature of Pul-SH3 was determined to be about 52 °C. The enzyme was able to hydrolyze pullulan, soluble starch, potato starch, and rice flour. It was exceptionally tolerant in the pH range of 4-11, exhibiting maximum activity at pH 8.5 and more than 60% of the activity in the pH range of 5-11. Being a detergent-tolerant pullulanase, Pul-SH3 retained 99, 89, and 54% of its activity at 10% concentration of Triton-X100, Tween 20, and SDS, respectively. The enzyme also exhibited an activity of 80.4 and 93.7% in the presence of two commercial detergents, Rika (7.5% v/v) and Fadisheh (2.5% w/v), respectively. The enzyme was even able to remain active by 54.5 and 85% after 10-day holding with the commercial detergents. Thermal stability of the enzyme could w on silica. Pul-SH3 with several industrially important characteristics seems desirable for cold hydrolysis of starch. PMID:26349928

  8. The first structure of a cold-adapted superoxide dismutase (SOD): biochemical and structural characterization of iron SOD from Aliivibrio salmonicida

    PubMed Central

    Pedersen, Hege Lynum; Willassen, Nils Peder; Leiros, Ingar

    2009-01-01

    Superoxide dismutases (SODs) are metalloenzymes that catalyse the dismutation of the superoxide radical anion into O2 and H2O2 in a two-step reaction. The crystal structure of the iron superoxide dismutase from the cold-adapted and fish-pathogenic bacterium Aliivibrio salmonicida (asFeSOD) has been determined and refined to 1.7 Å resolution. The protein has been characterized and compared with the closely related homologous iron superoxide dismutase from the mesophilic Escherichia coli (ecFeSOD) in an attempt to rationalize its environmental adaptation. ecFeSOD shares 75% identity with asFeSOD. Compared with the mesophilic FeSOD, the psychrophilic FeSOD has distinct temperature differences in residual activity and thermostability that do not seem to be related to structural differences such as intramolecular or intermolecular ion bonds, hydrogen bonds or cavity sizes. However, an increased net negative charge on the surface of asFeSOD may explain its lower thermostability com­­pared with ecFeSOD. Activity measurements and differential scanning calori­metry measurements revealed that the psychrophilic asFeSOD had a thermostability that was significantly higher than the optimal growth temperature of the host organism. PMID:19193992

  9. A novel cold-adapted and highly salt-tolerant esterase from Alkalibacterium sp. SL3 from the sediment of a soda lake.

    PubMed

    Wang, Guozeng; Wang, Qiaohuang; Lin, Xianju; Ng, Tzi Bun; Yan, Renxiang; Lin, Juan; Ye, Xiuyun

    2016-01-01

    A novel esterase gene (estSL3) was cloned from the Alkalibacterium sp. SL3, which was isolated from the sediment of soda lake Dabusu. The 636-bp full-length gene encodes a polypeptide of 211 amino acid residues that is closely related with putative GDSL family lipases from Alkalibacterium and Enterococcus. The gene was successfully expressed in E. coli, and the recombinant protein (rEstSL3) was purified to electrophoretic homogeneity and characterized. rEstSL3 exhibited the highest activity towards pNP-acetate and had no activity towards pNP-esters with acyl chains longer than C8. The enzyme was highly cold-adapted, showing an apparent temperature optimum of 30 °C and remaining approximately 70% of the activity at 0 °C. It was active and stable over the pH range from 7 to 10, and highly salt-tolerant up to 5 M NaCl. Moreover, rEstSL3 was strongly resistant to most tested metal ions, chemical reagents, detergents and organic solvents. Amino acid composition analysis indicated that EstSL3 had fewer proline residues, hydrogen bonds and salt bridges than mesophilic and thermophilic counterparts, but more acidic amino acids and less hydrophobic amino acids when compared with other salt-tolerant esterases. The cold active, salt-tolerant and chemical-resistant properties make it a promising enzyme for basic research and industrial applications.

  10. A novel cold-adapted and highly salt-tolerant esterase from Alkalibacterium sp. SL3 from the sediment of a soda lake

    PubMed Central

    Wang, Guozeng; Wang, Qiaohuang; Lin, Xianju; Bun Ng, Tzi; Yan, Renxiang; Lin, Juan; Ye, Xiuyun

    2016-01-01

    A novel esterase gene (estSL3) was cloned from the Alkalibacterium sp. SL3, which was isolated from the sediment of soda lake Dabusu. The 636-bp full-length gene encodes a polypeptide of 211 amino acid residues that is closely related with putative GDSL family lipases from Alkalibacterium and Enterococcus. The gene was successfully expressed in E. coli, and the recombinant protein (rEstSL3) was purified to electrophoretic homogeneity and characterized. rEstSL3 exhibited the highest activity towards pNP-acetate and had no activity towards pNP-esters with acyl chains longer than C8. The enzyme was highly cold-adapted, showing an apparent temperature optimum of 30 °C and remaining approximately 70% of the activity at 0 °C. It was active and stable over the pH range from 7 to 10, and highly salt-tolerant up to 5 M NaCl. Moreover, rEstSL3 was strongly resistant to most tested metal ions, chemical reagents, detergents and organic solvents. Amino acid composition analysis indicated that EstSL3 had fewer proline residues, hydrogen bonds and salt bridges than mesophilic and thermophilic counterparts, but more acidic amino acids and less hydrophobic amino acids when compared with other salt-tolerant esterases. The cold active, salt-tolerant and chemical-resistant properties make it a promising enzyme for basic research and industrial applications. PMID:26915906

  11. Gene cloning and catalytic characterization of cold-adapted lipase of Photobacterium sp. MA1-3 isolated from blood clam.

    PubMed

    Kim, Young Ok; Khosasih, Vivia; Nam, Bo-Hye; Lee, Sang-Jun; Suwanto, Antonius; Kim, Hyung Kwoun

    2012-12-01

    A lipase-producing Photobacterium strain (MA1-3) was isolated from the intestine of a blood clam caught at Namhae, Korea. The lipase gene was cloned by shotgun cloning and encoded 340 amino acids with a molecular mass of 38,015 Da. It had a very low sequence identity with other bacterial lipases, with the exception of that of Photobacterium lipolyticum M37 (83.2%). The MA1-3 lipase was produced in soluble form when Escherichia coli cells harboring the gene were cultured at 18°C. Its optimum temperature and pH were 45°C and pH 8.5, respectively. Its activation energy was calculated to be 2.69 kcal/mol, suggesting it to be a cold-adapted lipase. Its optimum temperature, temperature stability, and substrate specificity were quite different from those of M37 lipase, despite the considerable sequence similarities. Meanwhile, MA1-3 lipase performed a transesterification reaction using olive oil and various alcohols including methanol, ethanol, 1-propanol, and 1-butanol. In the presence of t-butanol as a co-solvent, this lipase produced biodiesel using methanol and plant or waste oils. The highest biodiesel conversion yield (73%) was achieved using waste soybean oil and methanol at a molar ratio of 1:5 after 12 h using 5 units of lipase. PMID:22841866

  12. Safety, immunogencity, and efficacy of a cold-adapted A/Ann Arbor/6/60 (H2N2) vaccine in mice and ferrets

    SciTech Connect

    Chen, Grace L.; Lamirande, Elaine W.; Jin Hong; Kemble, George; Subbarao, Kanta

    2010-03-01

    We studied the attenuation, immunogenicity and efficacy of the cold-adapted A/Ann Arbor/6/60 (AA ca) (H2N2) virus in mice and ferrets to evaluate its use in the event of an H2 influenza pandemic. The AA ca virus was restricted in replication in the respiratory tract of mice and ferrets. In mice, 2 doses of vaccine elicited a > 4-fold rise in hemagglutination-inhibition (HAI) titer and resulted in complete inhibition of viral replication following lethal homologous wild-type virus challenge. In ferrets, a single dose of the vaccine elicited a > 4-fold rise in HAI titer and conferred complete protection against homologous wild-type virus challenge in the upper respiratory tract. In both mice and ferrets, the AA ca virus provided significant protection from challenge with heterologous H2 virus challenge in the respiratory tract. The AA ca vaccine is safe, immunogenic, and efficacious against homologous and heterologous challenge in mice and ferrets, supporting the evaluation of this vaccine in clinical trials.

  13. Two cold-induced family 19 glycosyl hydrolases from cherimoya (Annona cherimola) fruit: an antifungal chitinase and a cold-adapted chitinase.

    PubMed

    Goñi, Oscar; Sanchez-Ballesta, María T; Merodio, Carmen; Escribano, María I

    2013-11-01

    Two cold-induced chitinases were isolated and purified from the mesocarp cherimoyas (Annona cherimola Mill.) and they were characterised as acidic endochitinases with a Mr of 24.79 and 47.77kDa (AChi24 and AChi48, respectively), both family 19 glycosyl hydrolases. These purified chitinases differed significantly in their biochemical and biophysical properties. While both enzymes had similar optimal acidic pH values, AChi24 was enzymatically active and stable at alkaline pH values, as well as displaying an optimal temperature of 45°C and moderate thermostability. Kinetic studies revealed a great catalytic efficiency of AChi24 for oligomeric and polymeric substrates. Conversely, AChi48 hydrolysis showed positive co-operativity that was associated to a mixture of different functional oligomeric states through weak transient protein interactions. The rise in the AChi48 kcat at increasing enzyme concentrations provided evidence of its oligomerisation. AChi48 chitinase was active and stable in a broad acidic pH range, and while it was relatively labile as temperatures increased, with an optimal temperature of 35°C, it retained about 50% of its maximal activity from 5 to 50°C. Thermodynamic characterisation reflected the high kcat of AChi48 and the remarkably lower ΔH(‡), ΔS(‡) and ΔG(‡) values at 5°C compared to AChi24, indicating that the hydrolytic activity of AChi48 was less thermodependent. In vitro functional studies revealed that AChi24 had a strong antifungal defence potential against Botrytis cinerea, whereas they displayed no cryoprotective or antifreeze activity. Hence, based on biochemical, thermodynamic and functional data, this study demonstrates that two acidic endochitinases are induced at low temperatures in a subtropical fruit, and that one of them acts in an oligomeric cold-adapted manner. PMID:23890591

  14. Characterization of a New Cold-Adapted and Salt-Activated Polysaccharide Lyase Family 7 Alginate Lyase from Pseudoalteromonas sp. SM0524

    PubMed Central

    Chen, Xiu-Lan; Dong, Sheng; Xu, Fei; Dong, Fang; Li, Ping-Yi; Zhang, Xi-Ying; Zhou, Bai-Cheng; Zhang, Yu-Zhong; Xie, Bin-Bin

    2016-01-01

    Marine bacterial alginate lyases play a role in marine alginate degradation and carbon cycling. Although a large number of alginate lyases have been characterized, reports on alginate lyases with special characteristics are still rather less. Here, a gene alyPM encoding an alginate lyase of polysaccharide lyase family 7 (PL7) was cloned from marine Pseudoalteromonas sp. SM0524 and expressed in Escherichia coli. AlyPM shows 41% sequence identity to characterized alginate lyases, indicating that AlyPM is a new PL7 enzyme. The optimal pH for AlyPM activity was 8.5. AlyPM showed the highest activity at 30°C and remained 19% of the highest activity at 5°C. AlyPM was unstable at temperatures above 30°C and had a low Tm of 37°C. These data indicate that AlyPM is a cold-adapted enzyme. Moreover, AlyPM is a salt-activated enzyme. AlyPM activity in 0.5–1.2 M NaCl was sixfolds higher than that in 0 M NaCl, probably caused by a significant increase in substrate affinity, because the Km of AlyPM in 0.5 M NaCl decreased more than 20-folds than that in 0 M NaCl. AlyPM preferably degraded polymannuronate and mainly released dimers and trimers. These data indicate that AlyPM is a new PL7 endo-alginate lyase with special characteristics. PMID:27486451

  15. Two cold-induced family 19 glycosyl hydrolases from cherimoya (Annona cherimola) fruit: an antifungal chitinase and a cold-adapted chitinase.

    PubMed

    Goñi, Oscar; Sanchez-Ballesta, María T; Merodio, Carmen; Escribano, María I

    2013-11-01

    Two cold-induced chitinases were isolated and purified from the mesocarp cherimoyas (Annona cherimola Mill.) and they were characterised as acidic endochitinases with a Mr of 24.79 and 47.77kDa (AChi24 and AChi48, respectively), both family 19 glycosyl hydrolases. These purified chitinases differed significantly in their biochemical and biophysical properties. While both enzymes had similar optimal acidic pH values, AChi24 was enzymatically active and stable at alkaline pH values, as well as displaying an optimal temperature of 45°C and moderate thermostability. Kinetic studies revealed a great catalytic efficiency of AChi24 for oligomeric and polymeric substrates. Conversely, AChi48 hydrolysis showed positive co-operativity that was associated to a mixture of different functional oligomeric states through weak transient protein interactions. The rise in the AChi48 kcat at increasing enzyme concentrations provided evidence of its oligomerisation. AChi48 chitinase was active and stable in a broad acidic pH range, and while it was relatively labile as temperatures increased, with an optimal temperature of 35°C, it retained about 50% of its maximal activity from 5 to 50°C. Thermodynamic characterisation reflected the high kcat of AChi48 and the remarkably lower ΔH(‡), ΔS(‡) and ΔG(‡) values at 5°C compared to AChi24, indicating that the hydrolytic activity of AChi48 was less thermodependent. In vitro functional studies revealed that AChi24 had a strong antifungal defence potential against Botrytis cinerea, whereas they displayed no cryoprotective or antifreeze activity. Hence, based on biochemical, thermodynamic and functional data, this study demonstrates that two acidic endochitinases are induced at low temperatures in a subtropical fruit, and that one of them acts in an oligomeric cold-adapted manner.

  16. Cold adaptation in marine organisms.

    PubMed

    Johnston, I A

    1990-01-30

    Animals from polar seas exhibit numerous so called resistance adaptations that serve to maintain homeostasis at low temperature and prevent lethal freezing injury. Specialization to temperatures at or below 0 degrees C is associated with an inability to survive at temperatures above 3-8 degrees C. Polar fish synthesize various types of glycoproteins or peptides to lower the freezing point of most extracellular fluid compartments in a non-colligative manner. Antifreeze production is seasonal in boreal species and is often initiated by environmental cues other than low temperature, particularly short day lengths. Most of the adaptations that enable intertidal invertebrates to survive freezing are associated with their ability to withstand ariel exposure. Unique adaptations for freezing avoidance include the synthesis of low molecular mass ice-nucleating proteins that control and induce extracellular ice-formation. Marine poikilotherms also exhibit a range of capacity adaptations that increase the rate of some physiological processes so as to partially compensate for the effects of low temperature. However, the rate of embryonic development in a diverse range of marine organisms shows no evidence of temperature compensation. This results in a significant lengthening of the time from fertilization to hatching in polar, relative to temperate, species. Some aspects of the physiology of polar marine species, such as low metabolic and slow growth rates, probably result from a combination of low temperature and other factors such as the highly seasonal nature of food supplies. Although neuromuscular function shows a partial capacity adaptation in Antarctic fish, maximum swimming speeds are lower than for temperate and tropical species, particularly for early stages in the life history. PMID:1969650

  17. [Phylogenetic diversity and cold-adaptive hydrolytic enzymes of culturable psychrophilic bacteria associated with sea ice from high latitude ocean, Artic].

    PubMed

    Yu, Yong; Li, Hui-Rong; Chen, Bo; Zeng, Yin-Xin; He, Jian-Feng

    2006-04-01

    showing 100% similarity each other are retrieved from the database, eleven from Antarctic seawater bacteria, three from Antarctic sea-ice bacteria, one from Spitzbergen sea-ice bacteria, two from Chukchi Sea sea-ice bacteria, two from Canadian Basin sea-ice bacteria (in this study) and one from uncultured bacterium clone PDA-OTU11 associated with the coral Pocillopora damicornis from the Great Barrier Reef. These may indicate that the physiological and geographic barriers appear to be permeable and some bacterial species can survive in different environment. The majority of the bacterial strains are able to secrete diversity cold-adaptive hydrolytic enzymes into the medium at 4 degrees C. The isolates that are able to degrade Tween-80, glutin, and starch account for, respectively, 62.6%, 51.4% and 40.5%.

  18. Live attenuated influenza vaccine--a review.

    PubMed

    Gasparini, R; Amicizia, D; Lai, P L; Panatto, D

    2011-09-01

    Owing to the variability of influenza viruses, vaccine composition needs to be up-dated annually. As many variables can influence their efficacy, vaccines are still considered "sub-optimal". Many studies have been carried out in recent years to improve vaccines. In particular, researchers and vaccine-producing corporations have focused on developing a live vaccine. Among the candidate vaccines, the strain developed by Maassab has recently been licensed in the USA and Europe, after extensive investigation. This vaccine is safe and well tolerated, and has shown very good genetic stability. Although vaccine recipients are able to spread the virus, transmission to close contacts is practically non-existent. Studies on cold-adapted attenuated influenza vaccines have demonstrated that such vaccines are effective, and sometimes more effective than inactivated influenza vaccines. Cold-adapted attenuated influenza vaccines therefore appear to be an important weapon against influenza. However, a more widespread use of these vaccines is to be recommended, especially in children, as the more acceptable way of administration can favour parental compliance.

  19. A live attenuated H7N7 candidate vaccine virus induces neutralizing antibody that confers protection from challenge in mice, ferrets and monkeys

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A live attenuated H7N7 candidate vaccine virus was generated by reverse genetics using the modified hemagglutinin (HA) and neuraminidase (NA) genes of HP A/Netherlands/219/03 (NL/03) (H7N7) wild-type (wt) virus and the six internal protein genes of the cold-adapted (ca) A/Ann Arbor/6/60 ca (AA ca) (...

  20. Molecular Basis of Live-Attenuated Influenza Virus

    PubMed Central

    He, Wen; Wang, Wei; Han, Huamin; Wang, Lei; Zhang, Ge; Gao, Bin

    2013-01-01

    Human influenza is a seasonal disease associated with significant morbidity and mortality. The most effective means for controlling infection and thereby reducing morbidity and mortality is vaccination with a three inactivated influenza virus strains mixture, or by intranasal administration of a group of three different live attenuated influenza vaccine strains. Comparing to the inactivated vaccine, the attenuated live viruses allow better elicitation of a long-lasting and broader immune (humoral and cellular) response that represents a naturally occurring transient infection. The cold-adapted (ca) influenza A/AA/6/60 (H2N2) (AA ca) virus is the backbone for the live attenuated trivalent seasonal influenza vaccine licensed in the United States. Similarly, the influenza A components of live-attenuated vaccines used in Russia have been prepared as reassortants of the cold-adapted (ca) H2N2 viruses, A/Leningrad/134/17/57-ca (Len/17) and A/Leningrad/134/47/57-ca (Len/47) along with virulent epidemic strains. However, the mechanism of temperature-sensitive attenuation is largely elusive. To understand how modification at genetic level of influenza virus would result in attenuation of human influenza virus A/PR/8/34 (H1N1,A/PR8), we investigated the involvement of key mutations in the PB1 and/or PB2 genes in attenuation of influenza virus in vitro and in vivo. We have demonstrated that a few of residues in PB1 and PB2 are critical for the phenotypes of live attenuated, temperature sensitive influenza viruses by minigenome assay and real-time PCR. The information of these mutation loci could be used for elucidation of mechanism of temperature-sensitive attenuation and as a new strategy for influenza vaccine development. PMID:23555969

  1. Principles underlying rational design of live attenuated influenza vaccines

    PubMed Central

    Jang, Yo Han

    2012-01-01

    Despite recent innovative advances in molecular virology and the developments of vaccines, influenza virus remains a serious burden for human health. Vaccination has been considered a primary countermeasure for prevention of influenza infection. Live attenuated influenza vaccines (LAIVs) are particularly attracting attention as an effective strategy due to several advantages over inactivated vaccines. Cold-adaptation, as a classical means for attenuating viral virulence, has been successfully used for generating safe and effective donor strains of LAIVs against seasonal epidemics and occasional pandemics. Recently, the advent of reverse genetics technique expedited a variety of rational strategies to broaden the pool of LAIVs. Considering the breadth of antigenic diversity of influenza virus, the pool of LAIVs is likely to equip us with better options for controlling influenza pandemics. With a brief reflection on classical attenuating strategies used at the initial stage of development of LAIVs, especially on the principles underlying the development of cold-adapted LAIVs, we further discuss and outline other attenuation strategies especially with respect to the rationales for attenuation, and their practicality for mass production. Finally, we propose important considerations for a rational vaccine design, which will provide us with practical guidelines for improving the safety and effectiveness of LAIVs. PMID:23596576

  2. Relative immunogenicity of the cold-adapted influenza virus A/Ann Arbor/6/60 (A/AA/6/60-ca), recombinants of A/AA/6/60-ca, and parental strains with similar surface antigens.

    PubMed Central

    Tannock, G A; Paul, J A; Barry, R D

    1984-01-01

    The immunogenicity of several cold-adapted (ca) viruses was compared in CSL mice with that of wild-type parental viruses with similar surface antigens, according to the vaccinating dose required to clear a challenge consisting of 10(4.5) 50% tissue culture infective doses of the wild-type virus. All ca viruses were less immunogenic than their wild-type parental strains by a factor of 10(1.3) to 10(3.4), probably due to the restricted capacity of ca viruses to replicate in the respiratory tracts of mice. However, their immunogenicity was considerably enhanced when two quite small doses were administered 3 weeks apart. The immunogenicity of ca viruses when administered in two doses and wild-type viruses when administered as a single dose varied according to their surface antigens. It was highest for viruses with the H2N2 A/Ann Arbor/6/60 and H3N2 A/Queensland/6/72 surface antigens and lowest for those with H1N1 A/HK/123/77 surface antigens. When two doses consisting of 10(5.0) 50% tissue culture infective doses of A/Ann Arbor/6/60-ca were administered at an interval of 3 weeks, solid immunity was induced against the wild-type A/Ann Arbor/6/60 parental virus, two heterologous H3N2 strains, and an H1N1 strain. PMID:6693167

  3. Assisted Living

    MedlinePlus

    ... but they don't need full-time nursing care. Some assisted living facilities are part of retirement ... change. Assisted living costs less than nursing home care. It is still fairly expensive. Older people or ...

  4. Live attenuated influenza viruses produced in a suspension process with avian AGE1.CR.pIX cells

    PubMed Central

    2012-01-01

    Background Current influenza vaccines are trivalent or quadrivalent inactivated split or subunit vaccines administered intramuscularly, or live attenuated influenza vaccines (LAIV) adapted to replicate at temperatures below body temperature and administered intranasally. Both vaccines are considered safe and efficient, but due to differences in specific properties may complement each other to ensure reliable vaccine coverage. By now, licensed LAIV are produced in embryonated chicken eggs. In the near future influenza vaccines for human use will also be available from adherent MDCK or Vero cell cultures, but a scalable suspension process may facilitate production and supply with vaccines. Results We evaluated the production of cold-adapted human influenza virus strains in the duck suspension cell line AGE1.CR.pIX using a chemically-defined medium. One cold-adapted A (H1N1) and one cold-adapted B virus strain was tested, as well as the reference strain A/PR/8/34 (H1N1). It is shown that a medium exchange is not required for infection and that maximum virus titers are obtained for 1 × 10-6 trypsin units per cell. 1 L bioreactor cultivations showed that 4 × 106 cells/mL can be infected without a cell density effect achieving titers of 1 × 108 virions/mL after 24 h. Conclusions Overall, this study demonstrates that AGE1.CR.pIX cells support replication of LAIV strains in a chemically-defined medium using a simple process without medium exchanges. Moreover, the process is fast with peak titers obtained 24 h post infection and easily scalable to industrial volumes as neither microcarriers nor medium replacements are required. PMID:23110398

  5. Assisted Living

    MedlinePlus

    ... it, too. Back to top What is the Cost for Assisted Living? Although assisted living costs less than nursing home care, it is still ... of services an older person chooses, the price costs can range from less than $25,000 a ...

  6. Role of influenza vaccine for healthy children in the US.

    PubMed

    Block, Stan L

    2004-01-01

    Influenza infection is associated with significant morbidity and mortality in adults, but the highest attack rates for influenza regularly occur in children, particularly those in preschool and elementary school. The consequences of influenza in this younger population - increased rate of hospitalization in those younger than 2 years of age and serious associated morbidity - have been underestimated. Children are also the critical link for spreading influenza in the community. Recent data suggest that mass influenza vaccination of healthy children would not only protect recipients, but also may reduce the burden of influenza throughout the community. During the past 3 decades, efforts to control influenza have focused on the use of an injectable trivalent inactivated vaccine (TIV) in high-risk persons. The vaccine is 'safe' and effective, but its acceptance and uptake by patients and healthcare providers have been modest at best. A new intranasal, live-attenuated, trivalent cold-adapted influenza virus vaccine (CAIV-T) [FluMist] is 'safe', well tolerated, immunogenic, and efficacious in preventing influenza illness in healthy children. Compared with TIV, CAIV-T is easier to administer and should be more readily acceptable, particularly for mass immunization campaigns. CAIV-T also induces a broader immune response and has demonstrated protection against at least three different variant influenza strains. This vaccine is particularly well suited for routine immunization of children and thus offers the potential for greatly improved control of influenza. However, the acquisition cost per single dose of FluMist for the 2003-4 season ( approximate, equals 46 US dollars) significantly hampered its uptake both by practitioners and by managed care organizations, even despite a later approximate, equals 25 US dollars rebate offer. For the 2004-5 season, CAIV-T is likely to be only modestly more expensive (average wholesale price: 16.50 US dollars for non-returnable doses

  7. Assisted Living

    MedlinePlus

    ... Recreational activities Security Transportation How to Choose a Facility A good match between a facility and a resident's needs depends as much on the philosophy and services of the assisted living facility as it does on the quality of care. ...

  8. Bachelor Living

    ERIC Educational Resources Information Center

    Germer, Sondra

    1974-01-01

    Male high school students in a Bachelor Living Class observed methods of child care including bottle feeding, spoon feeding, changing diapers, and method of holding. The purpose was for the students to grasp a better understanding of child development. (EK)

  9. Healthy Living

    MedlinePlus

    ... Environment Kids Health Kids Environment Kids Health Topics Environment & Health Healthy Living Pollution Reduce, Reuse, Recycle Science – How It Works The Natural World Games Brainteasers Puzzles Riddles Songs Activities Be ...

  10. Living Laboratories

    ERIC Educational Resources Information Center

    Mules, B. R.

    1976-01-01

    Presented is a review of various methods of keeping live animals, including scorpions, spiders, crabs, crayfish, shrimp, ants, fish, mice, and birds, as well as plants as a school science project/display. (SL)

  11. Assisted Living

    MedlinePlus

    ... premises. Adult foster care has the advantages of maintaining frail older adults in a more home-like ... pay to live in these communities, though some facilities have beds for skilled care that are funded ...

  12. Ontology-based combinatorial comparative analysis of adverse events associated with killed and live influenza vaccines.

    PubMed

    Sarntivijai, Sirarat; Xiang, Zuoshuang; Shedden, Kerby A; Markel, Howard; Omenn, Gilbert S; Athey, Brian D; He, Yongqun

    2012-01-01

    Vaccine adverse events (VAEs) are adverse bodily changes occurring after vaccination. Understanding the adverse event (AE) profiles is a crucial step to identify serious AEs. Two different types of seasonal influenza vaccines have been used on the market: trivalent (killed) inactivated influenza vaccine (TIV) and trivalent live attenuated influenza vaccine (LAIV). Different adverse event profiles induced by these two groups of seasonal influenza vaccines were studied based on the data drawn from the CDC Vaccine Adverse Event Report System (VAERS). Extracted from VAERS were 37,621 AE reports for four TIVs (Afluria, Fluarix, Fluvirin, and Fluzone) and 3,707 AE reports for the only LAIV (FluMist). The AE report data were analyzed by a novel combinatorial, ontology-based detection of AE method (CODAE). CODAE detects AEs using Proportional Reporting Ratio (PRR), Chi-square significance test, and base level filtration, and groups identified AEs by ontology-based hierarchical classification. In total, 48 TIV-enriched and 68 LAIV-enriched AEs were identified (PRR>2, Chi-square score >4, and the number of cases >0.2% of total reports). These AE terms were classified using the Ontology of Adverse Events (OAE), MedDRA, and SNOMED-CT. The OAE method provided better classification results than the two other methods. Thirteen out of 48 TIV-enriched AEs were related to neurological and muscular processing such as paralysis, movement disorders, and muscular weakness. In contrast, 15 out of 68 LAIV-enriched AEs were associated with inflammatory response and respiratory system disorders. There were evidences of two severe adverse events (Guillain-Barre Syndrome and paralysis) present in TIV. Although these severe adverse events were at low incidence rate, they were found to be more significantly enriched in TIV-vaccinated patients than LAIV-vaccinated patients. Therefore, our novel combinatorial bioinformatics analysis discovered that LAIV had lower chance of inducing these two

  13. Independent Living.

    ERIC Educational Resources Information Center

    Nathanson, Jeanne H., Ed.

    1994-01-01

    This issue of "OSERS" addresses the subject of independent living of individuals with disabilities. The issue includes a message from Judith E. Heumann, the Assistant Secretary of the Office of Special Education and Rehabilitative Services (OSERS), and 10 papers. Papers have the following titles and authors: "Changes in the Rehabilitation Act of…

  14. Healthy Living

    MedlinePlus

    ... health. Some you cannot control, such as your genetic makeup or your age. But you can make changes to your lifestyle. By taking steps toward healthy living, you can help reduce your risk of heart disease, cancer, stroke and other serious diseases: Get ...

  15. Retiring Lives

    ERIC Educational Resources Information Center

    Carnell, Eileen, Ed.; Lodge, Caroline, Ed.

    2009-01-01

    "Retiring Lives" presents fourteen personal real life stories from people at various stages of retiring. Each author recounts their own story about retiring, bringing together many aspects of the experiences: the social, psychological and practical. These inspirational and illustrated stories will encourage the reader to hold up these experiences…

  16. Outdoor Living.

    ERIC Educational Resources Information Center

    Cotter, Kathy

    Course objectives and learning activities are contained in this curriculum guide for a 16-week home economics course which teaches cooking and sewing skills applicable to outdoor living. The course goals include increasing male enrollment in the home economics program, developing students' self-confidence and ability to work in groups, and…

  17. Living History

    ERIC Educational Resources Information Center

    Walsh, Mark

    2005-01-01

    John Tinker and Mary Beth Tinker are back in a classroom in their hometown, once again wearing black armbands and drawing attention to a war. Now in their 50s, the siblings are living symbols of constitutional rights for secondary school students. In 1965, they and a handful of others were suspended for wearing black armbands to their public…

  18. Characterization of an extremely thermostable but cold-adaptive β-galactosidase from the hyperthermophilic archaeon Pyrococcus furiosus for use as a recombinant aggregation for batch lactose degradation at high temperature.

    PubMed

    Dong, Qing; Yan, Xufan; Zheng, Minhui; Yang, Ziwen

    2014-06-01

    β-Galactosidase (lactase), which catalyzes the hydrolysis of lactose into glucose and galactose, is one of the most important enzymes used in dairy processing. In this study, a gene that encoded an extremely thermostable β-galactosidase from Pyrococcus furiosus (Pflactase) was cloned and expressed in Escherichia coli BL21. The recombinant enzyme was purified by heat treatment and Ni-NTA affinity chromatography. The enzyme displayed optimal activity at 90°C and pH 7.0 in phosphate buffer. The specific activity of the recombinant enzyme on o-nitrophenyl-β-d-galactopyranoside was 10.2 U/mg at 0°C and 130.0dU/mg at 90°C. The half-lives of the enzyme were 31423.4, 8168.3, 4017.7, 547.4, 309.6, and 203.5 min at 70°C, 80°C, 85°C, 90°C, 95°C, and 100°C, respectively. The recombinant enzyme exhibited both β-galactosidase and β-glucosidase activity. The active inclusion bodies of β-galactosidase were easily isolated by nonionic detergent treatment and directly used for lactose conversion in a repetitive batch mode. More than 54% (90°C) or 88% (10°C) of the original enzyme activity was retained after 10 conversion cycles under optimum conditions. These results suggest that the recombinant thermostable β-galactosidase may be suitable for the hydrolysis of lactose in milk processing.

  19. ISS Live!

    NASA Technical Reports Server (NTRS)

    Price, Jennifer; Harris, Philip; Hochstetler, Bruce; Guerra, Mark; Mendez, Israel; Healy, Matthew; Khan, Ahmed

    2013-01-01

    International Space Station Live! (ISSLive!) is a Web application that uses a proprietary commercial technology called Lightstreamer to push data across the Internet using the standard http port (port 80). ISSLive! uses the push technology to display real-time telemetry and mission timeline data from the space station in any common Web browser or Internet- enabled mobile device. ISSLive! is designed to fill a unique niche in the education and outreach areas by providing access to real-time space station data without a physical presence in the mission control center. The technology conforms to Internet standards, supports the throughput needed for real-time space station data, and is flexible enough to work on a large number of Internet-enabled devices. ISSLive! consists of two custom components: (1) a series of data adapters that resides server-side in the mission control center at Johnson Space Center, and (2) a set of public html that renders the data pushed from the data adapters. A third component, the Lightstreamer server, is commercially available from a third party and acts as an intermediary between custom components (1) and (2). Lightstreamer also provides proprietary software libraries that are required to use the custom components. At the time of this reporting, this is the first usage of Web-based, push streaming technology in the aerospace industry.

  20. Live Virus Smallpox Vaccine

    MedlinePlus

    ... Index SMALLPOX FACT SHEET The Live Virus Smallpox Vaccine The vaccinia virus is the "live virus" used ... cannot cause smallpox. What is a "live virus" vaccine? A "live virus" vaccine is a vaccine that ...

  1. Living Nanomachines

    NASA Astrophysics Data System (ADS)

    Carlier, M.-F.; Helfer, E.; Wade, R.; Haraux, F.

    The living cell is a kind of factory on the microscopic scale, in which an assembly of modular machines carries out, in a spatially and temporally coordinated way, a whole range of activities internal to the cell, including the synthesis of substances essential to its survival, intracellular traffic, waste disposal, and cell division, but also activities related to intercellular communication and exchanges with the outside world, i.e., the ability of the cell to change shape, to move within a tissue, or to organise its own defence against attack by pathogens, injury, and so on. These nanomachines are made up of macromolecular assemblies with varying degrees of complexity, forged by evolution, within which work is done as a result of changes in interactions between proteins, or between proteins and nucleic acids, or between proteins and membrane components. All these cell components measure a few nanometers across, so the mechanical activity of these nanomachines all happens on the nanometric scale. The directional nature of the work carried out by biological nanomachines is associated with a dissipation of energy. As examples of protein assemblies, one could mention the proteasome, which is responsible for the degradation of proteins, and linear molecular motors such as actomyosin, responsible for muscle contraction, the dynein-microtubule system, responsible for flagellar motility, and the kinesin-microtubule system, responsible for transport of vesicles, which transform chemical energy into motion. Nucleic acid-protein assemblies include the ribosome, responsible for synthesising proteins, polymerases, helicases, elongation factors, and the machinery of DNA replication and repair; the mitotic spindle is an integrated system involving several of these activities which drive chromosome segregation. The machinery coupling membranes and proteins includes systems involved in the energy metabolism, such as the ATP synthase rotary motor, signalling cascades, endocytosis

  2. Engineering a substrate-specific cold-adapted subtilisin.

    PubMed

    Tindbaek, Nikolaj; Svendsen, Allan; Oestergaard, Peter Rahbek; Draborg, Henriette

    2004-02-01

    One region predicted to be highly flexible for a psychrophilic enzyme, TA39 subtilisin (S39), was transferred in silico to the mesophilic subtilisin, savinase (EC 3.4.21.62), from Bacillus lentus (clausii). The engineered hybrid and savinase were initially investigated by molecular dynamic simulations at 300 K to show binding region and global flexibility. The predicted S39 region consists of 12 residues, which due to homology between the subtilisins, results in a total change of eight residues. By site-directed modifications, the region was transferred to the binding region of savinase, thus a savinase-S39 hybrid, named H5, was constructed. The designed hybrid showed the same temperature optimum and pH profile as savinase, but H5 had higher specific activity on the synthetic substrate N-succinyl-L-Ala-L-Ala-L-Pro-L-Phe-p-nitroanilide (AAPF) at all temperatures measured and, at the same time, H5 showed a decrease in thermostability. The H5 hybrid showed broader substrate specificity, measured at room temperature, due to an increase in catalytic efficiency on AAPF, AAPA and FAAF compared with savinase (N-succinyl-XXXX-pNA; XXXX = AAPF, AAPA and FAAF). The H5 hybrid showed increased activity at low temperature, increased binding region and global flexibility, as investigated by molecular dynamic simulations, and global destabilization from differential scanning calorimetry measurements. These psychrophilic characteristics indicated an increase in binding site flexibility, probably due to the modifications P129S, S130G, P131E, and thus we show that it is possible to increase low temperature activity and global flexibility by engineered flexibility in the binding region. PMID:15047911

  3. Living with Sarcoidosis

    MedlinePlus

    ... page from the NHLBI on Twitter. Living With Sarcoidosis Sarcoidosis has no cure, but you can take ... Content: NEXT >> Featured Video Living With and Managing Sarcoidosis 05/18/2011 This video—presented by the ...

  4. Administration for Community Living

    MedlinePlus

    ... Information for Current Grantees About ACL Organization Why Community Living? Authorizing Statutes Budget Mandatory Grant Allocations Strategic ... Final Rule Get ACL Updates OAA Reauthorization Why Community Living? FEATURES #InclusionWorks IL Final Rule Get ACL ...

  5. The transcriptome of the bowhead whale Balaena mysticetus reveals adaptations of the longest-lived mammal.

    PubMed

    Seim, Inge; Ma, Siming; Zhou, Xuming; Gerashchenko, Maxim V; Lee, Sang-Goo; Suydam, Robert; George, John C; Bickham, John W; Gladyshev, Vadim N

    2014-10-01

    Mammals vary dramatically in lifespan, by at least two-orders of magnitude, but the molecular basis for this difference remains largely unknown. The bowhead whale Balaena mysticetus is the longest-lived mammal known, with an estimated maximal lifespan in excess of two hundred years. It is also one of the two largest animals and the most cold-adapted baleen whale species. Here, we report the first genome-wide gene expression analyses of the bowhead whale, based on the de novo assembly of its transcriptome. Bowhead whale or cetacean-specific changes in gene expression were identified in the liver, kidney and heart, and complemented with analyses of positively selected genes. Changes associated with altered insulin signaling and other gene expression patterns could help explain the remarkable longevity of bowhead whales as well as their adaptation to a lipid-rich diet. The data also reveal parallels in candidate longevity adaptations of the bowhead whale, naked mole rat and Brandt's bat. The bowhead whale transcriptome is a valuable resource for the study of this remarkable animal, including the evolution of longevity and its important correlates such as resistance to cancer and other diseases.

  6. The transcriptome of the bowhead whale Balaena mysticetus reveals adaptations of the longest-lived mammal

    PubMed Central

    Seim, Inge; Ma, Siming; Zhou, Xuming; Gerashchenko, Maxim V.; Lee, Sang-Goo; Suydam, Robert; George, John C.; Bickham, John W.; Gladyshev, Vadim N.

    2014-01-01

    Mammals vary dramatically in lifespan, by at least two-orders of magnitude, but the molecular basis for this difference remains largely unknown. The bowhead whale Balaena mysticetus is the longest-lived mammal known, with an estimated maximal lifespan in excess of two hundred years. It is also one of the two largest animals and the most cold-adapted baleen whale species. Here, we report the first genome-wide gene expression analyses of the bowhead whale, based on the de novo assembly of its transcriptome. Bowhead whale or cetacean-specific changes in gene expression were identified in the liver, kidney and heart, and complemented with analyses of positively selected genes. Changes associated with altered insulin signaling and other gene expression patterns could help explain the remarkable longevity of bowhead whales as well as their adaptation to a lipid-rich diet. The data also reveal parallels in candidate longevity adaptations of the bowhead whale, naked mole rat and Brandt's bat. The bowhead whale transcriptome is a valuable resource for the study of this remarkable animal, including the evolution of longevity and its important correlates such as resistance to cancer and other diseases. PMID:25411232

  7. Field studies on the annual activity and the metabolic responses of a land snail population living in high altitude.

    PubMed

    Staikou, Alexandra; Tachtatzis, George; Feidantsis, Konstantinos; Michaelidis, Basile

    2016-01-01

    In the context of the metabolic cold adaptation hypothesis (MCA), we investigated a) the life and activity cycle characteristics and b) the metabolic responses of the endemic land snail species Cattania trizona olympica living at 1100m altitude in Olympus mountain (Greece). Field observations on the annual activity cycle of C. trizona olympica revealed that snails' activity was restricted mainly between the end of May and September, when the higher temperatures were recorded, while first matings were recorded in July and the last ones in mid September indicating a restricted favorable time period for reproduction. The activities of enzymes of intermediate metabolism showed a periodic seasonal pattern of change which seems to be closely related to the pattern of annual changes of air temperature and most of them exhibited higher activities during the coldest and warmest periods of the year. Moreover the data indicate a distinct differentiation of fuel oxidation during arousal and reproductive periods with lipid oxidation, apart from carbohydrates, contributing significantly to ATP turnover during reproductive activity. The higher enzymatic activities, determined in the tissues of C. trizona olympica than the corresponding ones determined in the tissues of the land snail species living at low altitudes, might indicate higher sensitivity of the intermediate metabolism and ATP turnover in C. trizona olympica to changes in environmental factors. Although the latter seems to be in line with the MCA hypothesis, it needs further investigation on metabolic rates to support it. PMID:26408810

  8. [Support in living].

    PubMed

    Krebs, H

    1993-06-01

    "Lebenshilfe" (help in living) is of crucial importance for the organization of live of persons with a handicap. This help in living, training and social integration, however, requires an image of the handicapped person that unrestrictedly recognizes her right to live and to be a human being with a handicap. But todays hedonistic and preference-utilitarian social trends make other, selecting value judgements. This article comments on these dubious ethical positions critically and develops a paradigmatic, positive point of view. This statement is partly based on the principles of 2 self-help organizations of the same name ("Lebenshilfe für geistig Behinderte") in Germany and in Austria.

  9. Living Willow Huts

    ERIC Educational Resources Information Center

    Keeler, Rusty

    2007-01-01

    Living Willow Huts are inexpensive to make, fun to plant, easy to grow, and make beautiful spaces for children. They involve planting dormant willow shoots in the ground and weaving them into shapes that will sprout and grow over time. People have been creating similar living architecture throughout the world for centuries in the forms of living…

  10. Is It Living?

    ERIC Educational Resources Information Center

    Keeley, Page

    2011-01-01

    The word "living" is commonly used throughout elementary science lessons that focus on the biological world. It is a word teachers often take for granted when teaching life science concepts. How similar the constructed meaning of a common word like "living" is to the meaning intended by the teacher or instructional materials depends on how a…

  11. Immunization with live nonpathogenic H5N3 duck influenza virus protects chickens against highly pathogenic H5N1 virus.

    PubMed

    Gambaryan, A S; Boravleva, E Y; Lomakina, N F; Kropotkina, E A; Gordeychuk, I V; Chvala, I A; Drygin, V V; Klenk, H-D; Matrosovich, M N

    2016-01-01

    Development of an effective, broadly-active and safe vaccine for protection of poultry from H5N1 highly pathogenic avian influenza viruses (HPAIVs) remains an important practical goal. In this study we used a low pathogenic wild aquatic bird virus isolate А/duck/Moscow/4182/2010 (H5N3) (dk/4182) as a live candidate vaccine. We compared this virus with four live 1:7 reassortant anti-H5N1 candidate vaccine viruses with modified hemagglutinin from either A/Vietnam/1203/04 (H5N1) or A/Kurgan/3/05 (H5N1) and the rest of the genes from either H2N2 cold-adapted master strain A/Leningrad/134/17/57 (rVN-Len and rKu-Len) or H6N2 virus A/gull/Moscow/3100/2006 (rVN-gull and rKu-gull). The viruses were tested in parallel for pathogenicity, immunogenicity and protective effectiveness in chickens using aerosol, intranasal and oral routes of immunization. All five viruses showed zero pathogenicity indexes in chickens. Viruses rVN-gull and rKu-gull were immunogenic and protective, but they were insufficiently attenuated and caused significant mortality of 1-day-old chickens. The viruses with cold-adapted backbones (rVN-Len and rKu-Len) were completely nonpathogenic, but they were significantly less immunogenic and provided lower protection against lethal challenge with HPAIV A/Chicken/Kurgan/3/05 (H5N1) as compared with three other vaccine candidates. Unlike other four viruses, dk/4182 was both safe and highly immunogenic in chickens of any age regardless of inoculation route. Single administration of 106 TCID50 of dk/4182 virus via drinking water provided complete protection of 30-days-old chickens from 100 LD50 of the challenge virus. Our results suggest that low pathogenic viruses of wild aquatic birds can be used as safe and effective live poultry vaccines against highly pathogenic avian viruses. PMID:27640442

  12. Engineering Living Functional Materials

    PubMed Central

    2016-01-01

    Natural materials, such as bone, integrate living cells composed of organic molecules together with inorganic components. This enables combinations of functionalities, such as mechanical strength and the ability to regenerate and remodel, which are not present in existing synthetic materials. Taking a cue from nature, we propose that engineered ‘living functional materials’ and ‘living materials synthesis platforms’ that incorporate both living systems and inorganic components could transform the performance and the manufacturing of materials. As a proof-of-concept, we recently demonstrated that synthetic gene circuits in Escherichia coli enabled biofilms to be both a functional material in its own right and a materials-synthesis platform. To demonstrate the former, we engineered E. coli biofilms into a chemical-inducer-responsive electrical switch. To demonstrate the latter, we engineered E. coli biofilms to dynamically organize biotic-abiotic materials across multiple length scales, template gold nanorods, gold nanowires, and metal/semiconductor heterostructures, and synthesize semiconductor nanoparticles (Chen, A. Y. et al. (2014) Synthesis and patterning of tunable multiscale materials with engineered cells. Nat. Mater.13, 515–523.). Thus, tools from synthetic biology, such as those for artificial gene regulation, can be used to engineer the spatiotemporal characteristics of living systems and to interface living systems with inorganic materials. Such hybrids can possess novel properties enabled by living cells while retaining desirable functionalities of inorganic systems. These systems, as living functional materials and as living materials foundries, would provide a radically different paradigm of materials performance and synthesis–materials possessing multifunctional, self-healing, adaptable, and evolvable properties that are created and organized in a distributed, bottom-up, autonomously assembled, and environmentally sustainable manner. PMID

  13. Living with Oxygen Therapy

    MedlinePlus

    ... page from the NHLBI on Twitter. Living With Oxygen Therapy Oxygen therapy helps many people function better and be ... chronic obstructive pulmonary disease) Although you may need oxygen therapy continuously or for long periods, it doesn' ...

  14. Live Your Life Well

    MedlinePlus

    ... about reasonable steps that if used consistently can increase your comfort and boost your ability to build a rewarding life. About the Live Your Life Well Campaign Mental Health America is the country's leading non-profit ...

  15. Living with Anemia

    MedlinePlus

    ... page from the NHLBI on Twitter. Living With Anemia Often, you can treat and control anemia. If ... by an inherited or chronic disease or trauma. Anemia and Children/Teens Infants and young children have ...

  16. Fluorescence Live Cell Imaging

    PubMed Central

    Ettinger, Andreas

    2014-01-01

    Fluorescence microscopy of live cells has become an integral part of modern cell biology. Fluorescent protein tags, live cell dyes, and other methods to fluorescently label proteins of interest provide a range of tools to investigate virtually any cellular process under the microscope. The two main experimental challenges in collecting meaningful live cell microscopy data are to minimize photodamage while retaining a useful signal-to-noise ratio, and to provide a suitable environment for cells or tissues to replicate physiological cell dynamics. This chapter aims to give a general overview on microscope design choices critical for fluorescence live cell imaging that apply to most fluorescence microscopy modalities, and on environmental control with a focus on mammalian tissue culture cells. In addition, we provide guidance on how to design and evaluate fluorescent protein constructs by spinning disk confocal microscopy. PMID:24974023

  17. Living with Hearing Loss

    MedlinePlus

    ... Issues Special Section: Focus on Communication Living with Hearing Loss Past Issues / Fall 2008 Table of Contents ... family, including dad Bob, have adapted to her hearing impairment. Photo courtesy of Stefan Radtke, www.stefanradtke. ...

  18. Living with Alopecia Areata

    MedlinePlus

    ... you wear a wig Sadness and depression Hopelessness Anger Embarrassment Guilt or self-blame that you somehow ... For siblings and other family members, shame and anger because the disease has also affected their lives ...

  19. Assisted Living Community Profile

    MedlinePlus

    ... News & Media News Releases Media Resources AHCA/NCAL Gazette Publications Social Media Resources & Publications Currently selected Assisted ... News & Media News Releases Media Resources AHCA/NCAL Gazette Publications Social Media Resources & Publications Assisted Living Studies ...

  20. Living with Pulmonary Hypertension

    MedlinePlus

    ... and Support Living with PH may cause fear, anxiety, depression, and stress. You may worry about your ... and friends also can help relieve stress and anxiety. Let your loved ones know how you feel ...

  1. Influenza Vaccine, Live Intranasal

    MedlinePlus

    ... the recombinant influenza vaccine (RIV). The nasal spray flu vaccine (live attenuated influenza vaccine or LAIV) should NOT ... to your doctor or pharmacist about the best flu vaccine option for you or your family.

  2. Living with Marfan Syndrome

    MedlinePlus

    ... live longer and enjoy a good quality of life. Many people who have Marfan syndrome and are ... tears and leaks blood. Aortic dissection is a life-threatening condition. The main symptom of aortic dissection ...

  3. Living with Paralysis

    MedlinePlus

    ... are available to answer your questions. Call toll-free 1-800-539-7309 Mon-Fri, 9am-5pm ... are people living with or impacted by paralysis. Free services and downloads > Paralysis Resource Guide Our free ...

  4. Implementation of new approaches for generating conventional reassortants for live attenuated influenza vaccine based on Russian master donor viruses.

    PubMed

    Shcherbik, Svetlana; Pearce, Nicholas; Kiseleva, Irina; Larionova, Natalie; Rudenko, Larisa; Xu, Xiyan; Wentworth, David E; Bousse, Tatiana

    2016-01-01

    Cold-adapted influenza strains A/Leningrad/134/17/57 (H2N2) and B/USSR/60/69, originally developed in Russia, have been reliable master donors of attenuation for preparing live attenuated influenza vaccines (LAIV). The classical strategy for generating LAIV reassortants is robust, but has some disadvantages. The generation of reassortants requires at least 3 passages under selective conditions after co-infection; each of these selective passages takes six days. Screening the reassortants for a genomic composition traditionally starts after a second limiting dilution cloning procedure, and the number of suitable reassortants is limited. We developed a new approach to shorten process of preparing LAIV seed viruses. Introducing the genotyping of reassortants by pyrosequencing and monitoring sequence integrity of surface antigens starting at the first selective passage allowed specific selection of suitable reassortants for the next cloning procedure and also eliminate one of the group selective passage in vaccine candidate generation. Homogeneity analysis confirmed that reducing the number of selective passages didn't affect the quality of LAIV seed viruses. Finally, the two-way hemagglutination inhibition test, implemented for all the final seed viruses, confirmed that any amino acid substitutions acquired by reassortants during egg propagation didn't affect antigenicity of the vaccine. Our new strategy reduces the time required to generate a vaccine and was used to generate seasonal LAIVs candidates for the 2012/2013, 2014/2015, and 2015/2016 seasons more rapidly. PMID:26519883

  5. Live biometric authenticity check

    NASA Astrophysics Data System (ADS)

    Szu, Harold H.; Hsu, Charles C.; Szu, Clifford; Wang, Shoujue

    2003-04-01

    This research defined the underpinning concepts of a system that was highly secure, yet was efficient and non-invasive enough for everyday use. The live biometric authenticity check augmented invariant fingerprints with variable live features offered superior security by combining physical characteristics of the user"s with a passcode (numerical PIN) or passphrase (a string of words), and might also easily be augmented with other biometric video imaging devices for the utmost security.

  6. Living-Cell Microarrays

    PubMed Central

    Yarmush, Martin L.; King, Kevin R.

    2011-01-01

    Living cells are remarkably complex. To unravel this complexity, living-cell assays have been developed that allow delivery of experimental stimuli and measurement of the resulting cellular responses. High-throughput adaptations of these assays, known as living-cell microarrays, which are based on microtiter plates, high-density spotting, microfabrication, and microfluidics technologies, are being developed for two general applications: (a) to screen large-scale chemical and genomic libraries and (b) to systematically investigate the local cellular microenvironment. These emerging experimental platforms offer exciting opportunities to rapidly identify genetic determinants of disease, to discover modulators of cellular function, and to probe the complex and dynamic relationships between cells and their local environment. PMID:19413510

  7. Living in the question.

    PubMed

    Flower, J

    1999-01-01

    We live in a fast moving-world. Business has accelerated to breathtaking speeds in the 1990s--and in the last few years the afterburner has really kicked in. The speed of change is overwhelming. Especially in health care, who has time to "live in the question?" We need to decide things quickly, get the decision out of the way, and move on, right? Maybe. Biology shows us that you can't plan ahead very far. New things come along that you don't even have a category for, and therefore you don't even see them. Things are going to happen that you literally have no notion are even possible. The key to succeeding in this environment? Don't plan ahead. Stay curious. Make small bets. Build organizational hothouses. Feed the seedlings that grow. The challenge is to remain curious, to live in the question, both personally and organizationally.

  8. Intestinal transplantation: living related.

    PubMed

    Pollard, S G

    1997-01-01

    The use of live donors in intestinal transplantation could potentially both reduce the severity of rejection responses against this highly immunogenic organ by better tissue matching and also reduce cold ischaemia times. These two advantages over cadaveric grafts could preserve mucosal integrity and reduce the risk of systemic sepsis from bacterial translocation. The disadvantages of live donation are the inherent risk to the donor and the compromise of using a shorter graft. Although only a handful of such cases have been performed, the success rate has been high and this is a therapeutic modality which should be explored further. PMID:9536535

  9. Cryopreservation of Living Organs

    NASA Astrophysics Data System (ADS)

    Tanasawa, Ichiro; Nagata, Shinichi; Kimura, Naohiro

    Cryopreservation is considered to be the most promising way of preserving living organs or tissues for a long period of time without casuing any damage to their biological functions. However, cryopreservation has been succeeded only for simple and small-size tissues such as spermatozoon, ovum, erythrocyte, bone marrow and cornea. Cryopreservation of more complex and large-scale organs are not yet succssful. The authors have attempted to establish a technique for cryopreservation of larger living organs. An experiment was carried out using daphnia (water flea). The optimum rates of freezing and thawing were determined together with the optimum selection of cryoprotectant. High recovery rate was achieved under these conditions.

  10. [Testing of apathogenic influenza virus H5N3 as a poultry live vaccine].

    PubMed

    Boravleva, E Y; Chvala, I A; Lomakina, N F; Repin, P I; Mudrak, N S; Rudenko, L G; Gambaryan, A S; Drygin, V V

    2015-01-01

    Four H5N2 experimental vaccine strains and the apathogenic wild duck H5N3 influenza virus A/duck/ Moscow/4182/2010 (dk/4182) were tested as a live poultry vaccine. Experimental strains had the hemagglutinin of the A/Vietnam/1203/04 strain lacking the polybasic HA cleavage site or the hemagglutinin from attenuated virus (Ku/ at) that was derived from the highly pathogenic influenza virus A/chicken/Kurgan/3/2005 (H5N1). The hemagglutinin of the Ku-at has the amino acid substitutions Asp54/Asn and Lys222/Thr in HA1 and Val48/Ile and Lys131/Thr in HA2, while maintaining the polybasic HA cleavage site at an invariable level. The other genes of these experimental strains were from the H2N2 cold-adapted master strain A/Leningrad/134/17/57 (VN-Len and Ku-Len) or from the apathogenic H6N2 virus A/gull/Moscow/3100/2006 (VN-Gull and Ku-Gull). A single immunization of mice with all tested strains elicited a high level of serum antibodies and provided complete protection against the challenge with the lethal dose of A/chicken/Kurgan/3/05. The pathogenicity indexes of the Ku-at and the other strains for chicken were virtually zero, whereas the index of the parent H5N1 virus A/chicken/Kurgan/3/2005 was 2.98. Intravenous, intranasal, and aerosol routes of vaccination were compared. It was shown that the strain dk/4182 was totally apathogenic for one-day-old chicken and provided complete protection against the highly pathogenic H5N1 virus. PMID:26665435

  11. Dementia and Assisted Living

    ERIC Educational Resources Information Center

    Hyde, Joan; Perez, Rosa; Forester, Brent

    2007-01-01

    Purpose: This article presents an overview of what is known about dementia services in assisted living settings and suggests areas for future research. Design and Methods: We undertook a search of Medline, the "Journals of Gerontology," and "The Gerontologist." We then organized publications dealing with the target subject into 10 topic areas and…

  12. Living with Cystic Fibrosis

    MedlinePlus

    ... from the NHLBI on Twitter. Living With Cystic Fibrosis If you or your child has cystic fibrosis (CF), you should learn as much as you ... about CF Care Centers, go to the Cystic Fibrosis Foundation's Care Center Network Web page. It's standard ...

  13. Microholography of Living Organisms.

    ERIC Educational Resources Information Center

    Solem, Johndale C.; Baldwin, George C.

    1982-01-01

    By using intense pulsed coherent x-ray sources it will be possible to obtain magnified three-dimensional images of living elementary biological structures at precisely defined instants. Discussed are sources/geometrics for x-ray holography, x-radiation interactions, factors affecting resolution, recording the hologram, high-intensity holography,…

  14. Test Pattern For Living.

    ERIC Educational Resources Information Center

    Johnson, Nicholas

    The lives of Americans today are ruled by interlocking corporations. These corporations together present only one kind of life as viable: the consumer's life. Television is their main means of presenting this view. One cannot choose something he does not know about, and many Americans are not sufficiently informed of the alternatives to make an…

  15. Living or Nonliving?

    ERIC Educational Resources Information Center

    Legaspi, Britt; Straits, William

    2011-01-01

    Categorizing organisms as living or nonliving things may seem to be intuitive by nature. Yet, it is regulated by scientific criteria. Students come to school with rules already in place. Their categorizing criteria have already been influenced by their personal experiences, also known as observations and inferences. They believe that all things…

  16. Living Systems Energy Module

    SciTech Connect

    1995-09-26

    The Living Systems Energy Module, renamed Voyage from the Sun, is a twenty-lesson curriculum designed to introduce students to the major ways in which energy is important in living systems. Voyage from the Sun tells the story of energy, describing its solar origins, how it is incorporated into living terrestrial systems through photosynthesis, how it flows from plants to herbivorous animals, and from herbivores to carnivores. A significant part of the unit is devoted to examining how humans use energy, and how human impact on natural habitats affects ecosystems. As students proceed through the unit, they read chapters of Voyage from the Sun, a comic book that describes the flow of energy in story form (Appendix A). During the course of the unit, an ``Energy Pyramid`` is erected in the classroom. This three-dimensional structure serves as a classroom exhibit, reminding students daily of the importance of energy and of the fragile nature of our living planet. Interactive activities teach students about adaptations that allow plants and animals to acquire, to use and to conserve energy. A complete list of curricular materials and copies of all activity sheets appear in Appendix B.

  17. You Live, You Learn

    ERIC Educational Resources Information Center

    Biesta, Gert

    2008-01-01

    The Learning Lives project, a four-year study into the learning biographies and trajectories of adults, was conducted by a team of researchers from the universities of Stirling, Exeter, Brighton and Leeds as part of the Teaching and Learning Research Programme (TLRP) of the Economic and Social Research Council, and has just been completed. Whereas…

  18. Design for Living

    ERIC Educational Resources Information Center

    Rosenblum, Todd

    2011-01-01

    Bringing a newborn home from the hospital can come with stress for any parent. Coming home with twins can be double the stress. This article shares the story of a couple faced with this situation 12 years ago with the birth of twins, one was born with complications. They lived in a Colonial until the twins were almost five years old, at which time…

  19. The Living Periodic Table

    ERIC Educational Resources Information Center

    Nahlik, Mary Schrodt

    2005-01-01

    To help make the abstract world of chemistry more concrete eighth-grade students, the author has them create a living periodic table that can be displayed in the classroom or hallway. This display includes information about the elements arranged in the traditional periodic table format, but also includes visual real-world representations of the…

  20. Moab's Living Room

    ERIC Educational Resources Information Center

    Berry, John N., III

    2007-01-01

    This article describes the Grand County Public Library (GCPL) which was awarded the 2007 Best Small Library in America, an award sponsored by "Library Journal" and the Bill & Melinda Gates Foundation. Some 4800 of Grand County, Utah's 8,826 people live in Moab and the rest in the adjacent Spanish Valley and environs. The locals are a sizable group…

  1. Live-cell imaging

    PubMed Central

    Cole, Richard

    2014-01-01

    It would be hard to argue that live-cell imaging has not changed our view of biology. The past 10 years have seen an explosion of interest in imaging cellular processes, down to the molecular level. There are now many advanced techniques being applied to live cell imaging. However, cellular health is often under appreciated. For many researchers, if the cell at the end of the experiment has not gone into apoptosis or is blebbed beyond recognition, than all is well. This is simply incorrect. There are many factors that need to be considered when performing live-cell imaging in order to maintain cellular health such as: imaging modality, media, temperature, humidity, PH, osmolality, and photon dose. The wavelength of illuminating light, and the total photon dose that the cells are exposed to, comprise two of the most important and controllable parameters of live-cell imaging. The lowest photon dose that achieves a measureable metric for the experimental question should be used, not the dose that produces cover photo quality images. This is paramount to ensure that the cellular processes being investigated are in their in vitro state and not shifted to an alternate pathway due to environmental stress. The timing of the mitosis is an ideal canary in the gold mine, in that any stress induced from the imaging will result in the increased length of mitosis, thus providing a control model for the current imagining conditions. PMID:25482523

  2. Living with Parkinson's

    MedlinePlus

    ... Tips from the Health Care Team Finding Resources Parkinson's HelpLine Learn More Educational Materials Do you want ... resources & more. Order Free Materials Today Living with Parkinson’s “Parkinson’s is a part of my life, but ...

  3. Loneliness and Living Arrangements

    ERIC Educational Resources Information Center

    Stancliffe, Roger J.; Lakin, K. Charlie; Doljanac, Robert; Byun, Soo-Yong; Taub, Sarah; Chiri, Giuseppina

    2007-01-01

    Adults with ID/DD live in increasingly small community settings, where the risk of loneliness may be greater. We examined self-reported loneliness among 1,002 individuals with ID/DD from 5 states in relation to community residence size, personal characteristics, social contact, and social climate. One third reported being lonely sometimes and one…

  4. Sheltered Living Conditions.

    ERIC Educational Resources Information Center

    Netherland's Central Society for Rehabilitation, The Hague.

    Resulting from a study conducted by the Advisory Housing Committee of the Dutch Society for Rehabilitation, the report describes housing conditions and possibilities for the physically handicapped in the Netherlands. Four categories of sheltered living conditions are described and analyzed: residential centers, supervised residential centers,…

  5. Family Living Supplement.

    ERIC Educational Resources Information Center

    Truitt, Debbie

    This family living supplement contains 125 supplemental ideas and strategies designed to help vocational home economics teachers increase student motivation and enrich the teaching process. Ideas and strategies are organized into seven sections. These are career planning, securing a job, and career success; managing financial resources, buying…

  6. Learning from Live Theater

    ERIC Educational Resources Information Center

    Greene, Jay P.; Hitt, Collin; Kraybill, Anne; Bogulski, Cari A.

    2015-01-01

    Culturally enriching field trips matter. They produce significant benefits for students on a variety of educational outcomes that schools and communities care about. This experiment on the effects of field trips to see live theater demonstrates that seeing plays is an effective way to teach academic content; increases student tolerance by…

  7. Living related liver transplantation.

    PubMed

    Makuuchi, M; Kawarazaki, H; Iwanaka, T; Kamada, N; Takayama, T; Kumon, M

    1992-01-01

    Liver transplantation from a brain death donor has not yet been accepted in Japan. The only alternative method at present is transplantation from a living donor. After the first successful living related liver transplantation was performed by Strong in Brisbane, Australia, Japanese hepatic and transplant surgeons also began to perform such operations. As of February 1991, 16 living related liver transplantations had already been performed in Japan, mainly for children with biliary atresia. Five of these patients subsequently died, however, our patient has survived more than 1 year, and she is presently leading a normal school life. The most important issue regarding living related liver transplantation is to ensure the donor's safety. For this purpose, we conducted a preoperative banking of the donor's own blood and plasma. In addition, a selective vascular occlusion was carried out to reduce blood loss during the resection of the liver. Intraoperative color Doppler ultrasonography was introduced for evaluating the circulation of the graft. By using this modality, the following three points were able to be accurately estimated in order to obtain optimal graft perfusion: 1) The most suitable position for the graft to be fixed to the abdominal wall, 2) whether or not the abdominal wall could be closed and 3) the indication for a ligation of the collateral veins to form a porto-systemic shunt. Thanks to these procedures, living related liver transplantations have now become an acceptable transplant method, however, a transplantation from a cadaver that is brain dead but still has a beating heart is still absolutely necessary for adult recipients. Therefore, in the future, both methods should be performed.

  8. Watersheds: where we live

    USGS Publications Warehouse

    Vandas, Stephen; Farrar, Frank

    1996-01-01

    We all live in a watershed. Animals and plants all live there with us. Everyone affects what happens in a watershed by how we treat the natural resources. So what is a watershed? It is the land area that drains water to a stream, river, lake, or ocean. Water travels over the Earth's surface across forest land, farm fields, pastures, suburban lawns, and city streets, or it seeps into the soil and makes its way to a stream as local ground water. Watersheds come in many different shapes and sizes. Some contain mountains and hills, and others are nearly flat. A watershed can be affected by many different activities and events. Construction of cities and towns, farming, logging, and the application and disposal of many garden and household chemicals can affect the quantity and quality of water flowing from a watershed.

  9. Living history biography

    SciTech Connect

    Puck, T.T.

    1994-11-15

    A living history biography is presented of Theodore T. Puck. This history is intimately involved with the progress towards mapping of the human genome through research at the forefront of molecular cytogenetics. A review of historical research aims such as human genetics studies based on somatic cells, isolation of mutants as genetic markers, complementation analysis, gene mapping and the measurement of mutation is presented. 37 refs., 4 figs.

  10. Improving assisted living care.

    PubMed

    Gregory, Nancy; Gesell, Sabina B; Widmer, Tom

    2007-01-01

    In the absence of a national measurement system, private vendors of satisfaction measurement and improvement services have played a crucial role in the quality movement in the assisted living industry. Survey responses from 175 resident-family dyads at 20 facilities were analyzed to identify priorities for service improvement from the customers' perspective. They include improving care provided by aides and management, meal service, and activities. Practical solutions for addressing these issues are presented.

  11. Interior view of former living porch, now living area extension, ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    Interior view of former living porch, now living area extension, facing east. - Albrook Air Force Station, Field Officer's Quarters, West side of Dargue Avenue Circle, Balboa, Former Panama Canal Zone, CZ

  12. Generation and Characterization of Live Attenuated Influenza A(H7N9) Candidate Vaccine Virus Based on Russian Donor of Attenuation

    PubMed Central

    Shcherbik, Svetlana; Pearce, Nicholas; Balish, Amanda; Jones, Joyce; Thor, Sharmi; Davis, Charles Todd; Pearce, Melissa; Tumpey, Terrence; Cureton, David; Chen, Li-Mei; Villanueva, Julie; Bousse, Tatiana L.

    2015-01-01

    Background Avian influenza A (H7N9) virus has emerged recently and continues to cause severe disease with a high mortality rate in humans prompting the development of candidate vaccine viruses. Live attenuated influenza vaccines (LAIV) are 6:2 reassortant viruses containing the HA and NA gene segments from wild type influenza viruses to induce protective immune responses and the six internal genes from Master Donor Viruses (MDV) to provide temperature sensitive, cold-adapted and attenuated phenotypes. Methodology/Principal Findings LAIV candidate A/Anhui/1/2013(H7N9)-CDC-LV7A (abbreviated as CDC-LV7A), based on the Russian MDV, A/Leningrad/134/17/57 (H2N2), was generated by classical reassortment in eggs and retained MDV temperature-sensitive and cold-adapted phenotypes. CDC-LV7A had two amino acid substitutions N123D and N149D (H7 numbering) in HA and one substitution T10I in NA. To evaluate the role of these mutations on the replication capacity of the reassortants in eggs, the recombinant viruses A(H7N9)RG-LV1 and A(H7N9)RG-LV2 were generated by reverse genetics. These changes did not alter virus antigenicity as ferret antiserum to CDC-LV7A vaccine candidate inhibited hemagglutination by homologous A(H7N9) virus efficiently. Safety studies in ferrets confirmed that CDC-LV7A was attenuated compared to wild-type A/Anhui/1/2013. In addition, the genetic stability of this vaccine candidate was examined in eggs and ferrets by monitoring sequence changes acquired during virus replication in the two host models. No changes in the viral genome were detected after five passages in eggs. However, after ten passages additional mutations were detected in the HA gene. The vaccine candidate was shown to be stable in the ferret model; post-vaccination sequence data analysis showed no changes in viruses collected in nasal washes present at day 5 or day 7. Conclusions/Significance Our data indicate that the A/Anhui/1/2013(H7N9)-CDC-LV7A reassortant virus is a safe and

  13. Live from the Arctic

    NASA Astrophysics Data System (ADS)

    Warnick, W. K.; Haines-Stiles, G.; Warburton, J.; Sunwood, K.

    2003-12-01

    For reasons of geography and geophysics, the poles of our planet, the Arctic and Antarctica, are places where climate change appears first: they are global canaries in the mine shaft. But while Antarctica (its penguins and ozone hole, for example) has been relatively well-documented in recent books, TV programs and journalism, the far North has received somewhat less attention. This project builds on and advances what has been done to date to share the people, places, and stories of the North with all Americans through multiple media, over several years. In a collaborative project between the Arctic Research Consortium of the United States (ARCUS) and PASSPORT TO KNOWLEDGE, Live from the Arctic will bring the Arctic environment to the public through a series of primetime broadcasts, live and taped programming, interactive virtual field trips, and webcasts. The five-year project will culminate during the 2007-2008 International Polar Year (IPY). Live from the Arctic will: A. Promote global understanding about the value and world -wide significance of the Arctic, B. Bring cutting-edge research to both non-formal and formal education communities, C. Provide opportunities for collaboration between arctic scientists, arctic communities, and the general public. Content will focus on the following four themes. 1. Pan-Arctic Changes and Impacts on Land (i.e. snow cover; permafrost; glaciers; hydrology; species composition, distribution, and abundance; subsistence harvesting) 2. Pan-Arctic Changes and Impacts in the Sea (i.e. salinity, temperature, currents, nutrients, sea ice, marine ecosystems (including people, marine mammals and fisheries) 3. Pan-Arctic Changes and Impacts in the Atmosphere (i.e. precipitation and evaporation; effects on humans and their communities) 4. Global Perspectives (i.e. effects on humans and communities, impacts to rest of the world) In The Earth is Faster Now, a recent collection of comments by members of indigenous arctic peoples, arctic

  14. Assisted living in 1489.

    PubMed

    Warren, Carol A B

    2012-10-01

    Much has been written about theories of aging in premodern times, but much less is known about theories of residential care. This article describes a handbook prescribing the building, staffing, and running of residences for seniors: The Gerontocomos, published in Italy in 1489 by Gabrielle Zerbi. The handbook describes in detail the physical environment and architecture, the qualifications of the administrator, and the hiring and training of assistants for what is essentially an Assisted Living Facility. Although ancient-to-Renaissance theories of aging differ widely from contemporary theories, some of the practical suggestions are not so different.

  15. Living with lightning

    SciTech Connect

    Lamarre, L.

    1994-01-01

    As many as 100 lightning flashes occur around the world each second. Electric utilities know well the impact of lightning in terms of dollars, lost productivity, and lives. EPRI research, which began with a study of lightning`s natural characteristics, has resulted in tools utilities can use to better track and prepare for thunderstorms. Recently the institute completed a series of tests using small rockets to trigger and direct lightning strikes. Now EPRI-sponsored researchers are developing a laser-based technology they believe will be able to guide thunderbolts safely to the ground and ultimately even to discharge thunderclouds.

  16. Living in Space

    NASA Technical Reports Server (NTRS)

    Brown, Ray (Editor)

    1993-01-01

    In this educational video from the 'Liftoff to Learning' series, astronauts from the STS-56 Mission (Ken Cockrell, Mike Foale, Ellen Ochoa, Steve Oswald, and Ken Cameron) explain and show through demonstrations how microgravity affects the way astronauts live onboard the Space Shuttle, and how these same daily habits or processes differ on Earth. A tour of the Space Shuttle is given, including the sleeping compartments, the kitchen area, the storage compartments, and the Waste Collection System (or WCS, as they call it). Daily habits (brushing teeth, shampooing hair and bathing, eating,...) are explained and actively illustrated, along with reasons of how these applications differ from their employment on Earth.

  17. Living donor nephrectomy.

    PubMed

    Jacobs, S C; Flowers, J L; Dunkin, B; Sklar, G N; Cho, E

    1999-03-01

    The need for more organs for kidney transplantation is increasing. Cadaver sources for these organs are stable, therefore living donation must increase if the need is to be met. Less perfect kidneys are now being transplanted. The pool of potential donors is being expanded. The process of kidney donation is being made easier in an effort to increase the number of donors. The donor work-up is being streamlined. Laparoscopic donor nephrectomy has been introduced, and appears to be promising as a technique of lessening donor pain and suffering, while maintaining excellent graft results.

  18. "Living versus Dead":

    PubMed Central

    Chakrabarti, Pratik

    2010-01-01

    Summary The Semple antirabies vaccine was developed by David Semple in India in 1911. Semple introduced a peculiarly British approach within the Pasteurian tradition by using carbolized dead virus. This article studies this unique phase of vaccine research between 1910 and 1935 to show that in the debates and laboratory experiments around the potency and safety of vaccines, categories like "living" and "dead" were often used as ideological and moral denominations. These abstract and ideological debates were crucial in defining the final configuration of the Semple vaccine, the most popular antirabies vaccine used globally, and also in shaping international vaccination policies. PMID:21037397

  19. Communication in Assisted Living*

    PubMed Central

    Williams, Kristine N.; Warren, Carol A.B.

    2009-01-01

    This study of communication in an Assisted Living Facility (ALF) focuses on staff’s interpretive frameworks and situational tactics for managing elderly residents. It is based on interviews with staff and residents in an ALF together with ethnographic fieldwork. As in other quasi-total institutions, staff members engage in control as well as care, monitoring residents for compliance with rules and directives. Residents, aware of the threat of being moved to a nursing home, also monitor their own behavior and cognition in comparison to other residents. Other communication issues include the infantilization of the elderly by staff, and the race, class, and ethnic prejudices of residents. PMID:20107612

  20. Living olefin polymerization processes

    DOEpatents

    Schrock, Richard R.; Baumann, Robert

    2003-08-26

    Processes for the living polymerization of olefin monomers with terminal carbon-carbon double bonds are disclosed. The processes employ initiators that include a metal atom and a ligand having two group 15 atoms and a group 16 atom or three group 15 atoms. The ligand is bonded to the metal atom through two anionic or covalent bonds and a dative bond. The initiators are particularly stable under reaction conditions in the absence of olefin monomer. The processes provide polymers having low polydispersities, especially block copolymers having low polydispersities. It is an additional advantage of these processes that, during block copolymer synthesis, a relatively small amount of homopolymer is formed.

  1. Living olefin polymerization processes

    DOEpatents

    Schrock, Richard R.; Baumann, Robert

    1999-01-01

    Processes for the living polymerization of olefin monomers with terminal carbon-carbon double bonds are disclosed. The processes employ initiators that include a metal atom and a ligand having two group 15 atoms and a group 16 atom or three group 15 atoms. The ligand is bonded to the metal atom through two anionic or covalent bonds and a dative bond. The initiators are particularly stable under reaction conditions in the absence of olefin monomer. The processes provide polymers having low polydispersities, especially block copolymers having low polydispersities. It is an additional advantage of these processes that, during block copolymer synthesis, a relatively small amount of homopolymer is formed.

  2. Living olefin polymerization processes

    DOEpatents

    Schrock, R.R.; Baumann, R.

    1999-03-30

    Processes for the living polymerization of olefin monomers with terminal carbon-carbon double bonds are disclosed. The processes employ initiators that include a metal atom and a ligand having two group 15 atoms and a group 16 atom or three group 15 atoms. The ligand is bonded to the metal atom through two anionic or covalent bonds and a dative bond. The initiators are particularly stable under reaction conditions in the absence of olefin monomer. The processes provide polymers having low polydispersities, especially block copolymers having low polydispersities. It is an additional advantage of these processes that, during block copolymer synthesis, a relatively small amount of homopolymer is formed.

  3. Microencapsulation Of Living Cells

    NASA Technical Reports Server (NTRS)

    Chang, Manchium; Kendall, James M.; Wang, Taylor G.

    1989-01-01

    In experimental technique, living cells and other biological materials encapsulated within submillimeter-diameter liquid-filled spheres. Sphere material biocompatible, tough, and compliant. Semipermeable, permitting relatively small molecules to move into and out of sphere core but preventing passage of large molecules. New technique promises to make such spherical capsules at high rates and in uniform, controllable sizes. Capsules injected into patient through ordinary hypodermic needle. Promising application for technique in treatment of diabetes. Also used to encapsulate pituitary cells and thyroid hormone adrenocortical cells for treatment of other hormonal disorders, to encapsulate other secreting cells for transplantation, and to package variety of pharmaceutical products and agricultural chemicals for controlled release.

  4. Living olefin polymerization processes

    DOEpatents

    Schrock, Richard R.; Bauman, Robert

    2006-11-14

    Processes for the living polymerization of olefin monomers with terminal carbon-carbon double bonds are disclosed. The processes employ initiators that include a metal atom and a ligand having two group 15 atoms and a group 16 atom or three group 15 atoms. The ligand is bonded to the metal atom through two anionic or covalent bonds and a dative bond. The initiators are particularly stable under reaction conditions in the absence of olefin monomer. The processes provide polymers having low polydispersities, especially block copolymers having low polydispersities. It is an additional advantage of these processes that, during block copolymer synthesis, a relatively small amount of homopolymer is formed.

  5. National Center for Assisted Living

    MedlinePlus

    ... indicators to help providers stay ahead of the curve. Key Dates NOV 11 Live! Q & A Session ... Assisted Living Studies Clinical Practice Guidelines Health Information Technology In-Service Training Tools Periodicals State Regulatory Review ...

  6. Living with Tuberous Sclerosis Complex

    MedlinePlus

    ... living and employment while others may deal with dating issues and reproductive concerns. Throughout their lives, those ... Overview Outreach Toolkit Government Action Team TS Alliance Online Support Community Facebook Twitter YouTube How to Make ...

  7. Healthy Living Slashes Cancer Risk

    MedlinePlus

    ... Services, or federal policy. More Health News on: Cancer Healthy Living Recent Health News Related MedlinePlus Health Topics Cancer Healthy Living About MedlinePlus Site Map FAQs Contact Us Get ...

  8. H7N9 Live Attenuated Influenza Vaccine Is Highly Immunogenic, Prevents Virus Replication, and Protects Against Severe Bronchopneumonia in Ferrets.

    PubMed

    de Jonge, Jørgen; Isakova-Sivak, Irina; van Dijken, Harry; Spijkers, Sanne; Mouthaan, Justin; de Jong, Rineke; Smolonogina, Tatiana; Roholl, Paul; Rudenko, Larisa

    2016-05-01

    Avian influenza viruses continue to cross the species barrier, and if such viruses become transmissible among humans, it would pose a great threat to public health. Since its emergence in China in 2013, H7N9 has caused considerable morbidity and mortality. In the absence of a universal influenza vaccine, preparedness includes development of subtype-specific vaccines. In this study, we developed and evaluated in ferrets an intranasal live attenuated influenza vaccine (LAIV) against H7N9 based on the A/Leningrad/134/17/57 (H2N2) cold-adapted master donor virus. We demonstrate that the LAIV is attenuated and safe in ferrets and induces high hemagglutination- and neuraminidase-inhibiting and virus-neutralizing titers. The antibodies against hemagglutinin were also cross-reactive with divergent H7 strains. To assess efficacy, we used an intratracheal challenge ferret model in which an acute severe viral pneumonia is induced that closely resembles viral pneumonia observed in severe human cases. A single- and two-dose strategy provided complete protection against severe pneumonia and prevented virus replication. The protective effect of the two-dose strategy appeared better than the single dose only on the microscopic level in the lungs. We observed, however, an increased lymphocytic infiltration after challenge in single-vaccinated animals and hypothesize that this a side effect of the model. PMID:26796670

  9. RACE AS LIVED EXPERIENCE

    PubMed Central

    Garcia, John A.; Sanchez, Gabriel R.; Sanchez-Youngman, Shannon; Vargas, Edward D.; Ybarra, Vickie D.

    2015-01-01

    A growing body of social science research has sought to conceptualize race as a multidimensional concept in which context, societal relations, and institutional dynamics are key components. Utilizing a specially designed survey, we develop and use multiple measures of race (skin color, ascribed race, and discrimination experiences) to capture race as “lived experience” and assess their impact on Latinos’ self-rated health status. We model these measures of race as a lived experience to test the explanatory power of race, both independently and as an integrated scale with categorical regression, scaling, and dimensional analyses. Our analyses show that our multiple measures of race have significant and negative effects on Latinos’ self-reported health. Skin color is a dominant factor that impacts self-reported health both directly and indirectly. We then advocate for the utilization of multiple measures of race, adding to those used in our analysis, and their application to other health and social outcomes. Our analysis provides important contributions across a wide range of health, illness, social, and political outcomes for communities of color. PMID:26681972

  10. Live From the Poles

    NASA Astrophysics Data System (ADS)

    Linder, C. A.; Kent, J.; Lippsett, L.

    2006-12-01

    International Polar Year presents an extraordinary opportunity to educate students and the public about science at the icy ends of the Earth. The goal of our proposal is to apply collaborative multimedia approaches to bring the story of four polar research expeditions to the general public and the classroom. The four expeditions (measurement of ice sheet dynamics in Greenland, a study of the McMurdo ecosystem over austral winter, installation of a buoy array in the Beaufort Gyre, and exploration of the Gakkel Ridge) were chosen based on their broad range of disciplines and relevance to the three primary IPY research emphasis areas defined by NSF. A science writer and a professional photographer will join each expedition and file dispatches for a daily Webcast. The posting will feature science updates, logistical challenges, team member profiles, and life at sea (or on the ice). The writer will also coordinate real-time phone patches from PIs in the field to audiences at the Museum of Science, Boston, the Smithsonian National Museum of Natural History, The Field Museum, Chicago, the Houston Museum of Natural Science, the Birch Aquarium, San Diego, the Pacific Science Center, Seattle, National Public Radio "Talk of the Nation: Science Friday," CBS News, and to student "reporters" writing for Scholastic Online. At the museums, the "Live from the Ice" interactive phone calls will be preceded by a background presentation by a scientist, who will also moderate the live discussion between the public and researchers in the field. A 20-30 minute satellite phone call will allow the public to ask the researchers questions about their research while it's happening. In addition to building and promoting an online experience, a museum exhibit featuring models of Arctic instruments and informative kiosks will be developed at the Woods Hole Oceanographic Institution Exhibit Center. Each of our partner museums will also provide a "leave-behind" component to continue to educate

  11. Country living with AIDS.

    PubMed

    Kletecka, C

    1998-12-01

    The number of people with HIV in rural areas is small, but it is growing. In rural areas, AIDS organizations must deal with a fragmented client base that is spread out over a large geographic area. The logistics and costs of maintaining these services are higher per person than in urban areas. A description of what it is like to live with HIV in rural Vermont is provided. Although most people in rural Vermont seem to accept the concept of AIDS, few have known someone with the disease. Specialized medical care is difficult to obtain because only one specialty clinic and three part-time sites exist in the state of Vermont. Many who require more complex care, with better treatment options, go out of state to receive it.

  12. Living With Semantic Dementia

    PubMed Central

    Sage, Karen; Wilkinson, Ray; Keady, John

    2014-01-01

    Semantic dementia is a variant of frontotemporal dementia and is a recently recognized diagnostic condition. There has been some research quantitatively examining care partner stress and burden in frontotemporal dementia. There are, however, few studies exploring the subjective experiences of family members caring for those with frontotemporal dementia. Increased knowledge of such experiences would allow service providers to tailor intervention, support, and information better. We used a case study design, with thematic narrative analysis applied to interview data, to describe the experiences of a wife and son caring for a husband/father with semantic dementia. Using this approach, we identified four themes: (a) living with routines, (b) policing and protecting, (c) making connections, and (d) being adaptive and flexible. Each of these themes were shared and extended, with the importance of routines in everyday life highlighted. The implications for policy, practice, and research are discussed. PMID:24532121

  13. Freezing of living cells

    SciTech Connect

    Mazur, P.

    1985-01-01

    It can be calculated that a living cell will survive more than 5000 years at -196/sup 0/C. This ability to essentially stop biological time has important implications in medicine and agriculture, and in biological research. In medicine the chief implications are in the banking of transplantable tissues and organs and in in vitro fertilization. In agriculture the applications stem in part from the role of frozen embryos in amplifying the number of calves produced by high quanlity cows. The problem is how can cells survive both the cooling to such very low temperatures and the return to normal temperatures. The answers involve fundamental characteristics of cells such as the permeability of their surface membranes to water and solutes. These characteristics determine whether or not cells undergo lethal internal ice formation and other response during freezing and thawing. 27 refs., 12 figs.

  14. Live cell NMR.

    PubMed

    Freedberg, Darón I; Selenko, Philipp

    2014-01-01

    Ever since scientists realized that cells are the basic building blocks of all life, they have been developing tools to look inside them to reveal the architectures and mechanisms that define their biological functions. Whereas "looking into cells" is typically said in reference to optical microscopy, high-resolution in-cell and on-cell nuclear magnetic resonance (NMR) spectroscopy is a powerful method that offers exciting new possibilities for structural and functional studies in and on live cells. In contrast to conventional imaging techniques, in- and on-cell NMR methods do not provide spatial information on cellular biomolecules. Instead, they enable atomic-resolution insights into the native cell states of proteins, nucleic acids, glycans, and lipids. Here we review recent advances and developments in both fields and discuss emerging concepts that have been delineated with these methods.

  15. Living with uncertainty

    SciTech Connect

    Rau, N.; Fong, C.C.; Grigg, C.H.; Silverstein, B.

    1994-11-01

    In the electric utility industry, only one thing can be guaranteed with absolute certainty: one lives and works with many unknowns. Thus, the industry has embraced probability methods to varying degrees over the last 25 years. These techniques aid decision makers in planning, operations, and maintenance by quantifying uncertainty. Examples include power system reliability, production costing simulation, and assessment of environmental factors. A series of brainstorming sessions was conducted by the Application of Probability Methods (APM) Subcommittee of the IEEE Power Engineering Society to identify research and development needs and to ask the question, ''where should we go from here '' The subcommittee examined areas of need in data development, applications, and methods for decision making. The purpose of this article is to share the thoughts of APM members with a broader audience to the findings and to invite comments and participation.

  16. Lives Worth Living: Religious Education and Social Movements

    ERIC Educational Resources Information Center

    Ayres, Jennifer R.

    2013-01-01

    When people of faith participate in movements for social change, how are their religious and moral identities formed, challenged, and transformed? Although they have explicit and tangible goals as they participate in advocacy, protest, and boycotts, religious social activists also, James Jasper argues, craft "lives worth living" (1997).…

  17. The living publication

    SciTech Connect

    Terwilliger, Thomas C.

    2012-06-04

    Within the ICSTI Insights Series we offer three articles on the 'living publication' that is already available to practitioners in the important field of crystal structure determination and analysis. While the specific examples are drawn from this particular field, we invite readers to draw parallels in their own fields of interest. The first article describes the present state of the crystallographic living publication, already recognized by an ALPSP (Association of Learned and Professional Society Publishers) Award for Publishing Innovation in 2006. The second article describes the potential impact on the record of science as greater post-publication analysis becomes more common within currently accepted data deposition practices, using processed diffraction data as the starting point. The third article outlines a vision for the further improvement of crystallographic structure reports within potentially achievable enhanced data deposition practices, based upon raw (unprocessed) diffraction data. The IUCr in its Commissions and Journals has for many years emphasized the importance of publications being accompanied by data and the interpretation of the data in terms of atomic models. This has been followed as policy by numerous other journals in the field and its cognate disciplines. This practice has been well served by databases and archiving institutions such as the Protein Data Bank (PDB), the Cambridge Crystallographic Data Centre (CCDC), and the Inorganic Crystal Structure Database (ICSD). Normally the models that are archived are interpretations of the data, consisting of atomic coordinates with their displacement parameters, along with processed diffraction data from X-ray, neutron or electron diffraction studies. In our current online age, a reader can not only consult the printed word, but can display and explore the results with molecular graphics software of exceptional quality. Furthermore, the routine availability of processed diffraction data allows

  18. Community Living Skills: Nutrition I.

    ERIC Educational Resources Information Center

    Kreps, Alice Roelofs; Dreith, Rita Vallero

    One of twenty course guides in the Community Living Skills Guide for the College for Living series, this document provides guidelines and workbook activities for the course, Nutrition I. The series of courses for developmentally disabled adults is intended to supplement residential programs and to aid in orienting institutionalized persons to…

  19. Living with Diabetic Heart Disease

    MedlinePlus

    ... Heart Disease » Living With Diabetic Heart Disease Explore Diabetic Heart Disease What Is... Causes Who Is at Risk Signs & Symptoms Diagnosis Treatments Prevention Living With Clinical Trials Links Related Topics Atherosclerosis Cardiomyopathy Coronary Heart Disease Heart Attack Heart Failure Send ...

  20. Framework for Healthful Living Education.

    ERIC Educational Resources Information Center

    North Carolina State Dept. of Public Instruction, Raleigh.

    The Healthful Living Education program promotes behaviors that contribute to a healthful lifestyle and improved quality of life for all students. The Framework for Healthy Living Education supports and reinforces the goals and objectives of its three major components: health education, physical education, and alcohol and other drugs. When the…

  1. Learning Lives and Alumni Voices

    ERIC Educational Resources Information Center

    Jacobs, Andrea; Leach, Camilla; Spencer, Stephanie

    2010-01-01

    Changes in governmental financial support are causing many would-be students to question the value of higher education or to consider attending a local university. Oral history testimonies provide a source for understanding the role that living, as well as working, within an academic community plays in the learning lives of its alumni. An…

  2. Community Living Skills Guide: Sexuality.

    ERIC Educational Resources Information Center

    Breen, Kathy

    One of twenty course guides in the Community Living Skills Guide for the College for Living series, this document provides guidelines and workbook activities for the course, Sexuality. The series of courses for developmentally disabled adults is intended to supplement residential programs and to aid in orienting institutionalized persons to…

  3. Generation of live attenuated novel influenza virus A/California/7/09 (H1N1) vaccines with high yield in embryonated chicken eggs.

    PubMed

    Chen, Zhongying; Wang, Weijia; Zhou, Helen; Suguitan, Amorsolo L; Shambaugh, Cindy; Kim, Lomi; Zhao, Jackie; Kemble, George; Jin, Hong

    2010-01-01

    Several live attenuated influenza virus A/California/7/09 (H1N1) (CA09) candidate vaccine variants that possess the hemagglutinin (HA) and neuraminidase (NA) gene segments from the CA09 virus and six internal protein gene segments from the cold-adapted influenza virus A/Ann Arbor/6/60 (H2N2) virus were generated by reverse genetics. The reassortant viruses replicated relatively poorly in embryonated chicken eggs. To improve virus growth in eggs, reassortants expressing the HA and NA of CA09 were passaged in MDCK cells and variants exhibiting large-plaque morphology were isolated. These variants replicated at levels approximately 10-fold higher than the rate of replication of the parental strains in embryonated chicken eggs. Sequence analysis indicated that single amino acid changes at positions 119, 153, 154, and 186 were responsible for the improved growth properties in MDCK cells and eggs. In addition, the introduction of a mutation at residue 155 that was previously shown to enhance the replication of a 1976 swine influenza virus also significantly improved the replication of the CA09 virus in eggs. Each variant was further evaluated for receptor binding preference, antigenicity, attenuation phenotype, and immunogenicity. Mutations at residues 153, 154, and 155 drastically reduced viral antigenicity, which made these mutants unsuitable as vaccine candidates. However, changes at residues 119 and 186 did not affect virus antigenicity or immunogenicity, justifying their inclusion in live attenuated vaccine candidates to protect against the currently circulating 2009 swine origin H1N1 viruses.

  4. Generation of Live Attenuated Novel Influenza Virus A/California/7/09 (H1N1) Vaccines with High Yield in Embryonated Chicken Eggs ▿

    PubMed Central

    Chen, Zhongying; Wang, Weijia; Zhou, Helen; Suguitan, Amorsolo L.; Shambaugh, Cindy; Kim, Lomi; Zhao, Jackie; Kemble, George; Jin, Hong

    2010-01-01

    Several live attenuated influenza virus A/California/7/09 (H1N1) (CA09) candidate vaccine variants that possess the hemagglutinin (HA) and neuraminidase (NA) gene segments from the CA09 virus and six internal protein gene segments from the cold-adapted influenza virus A/Ann Arbor/6/60 (H2N2) virus were generated by reverse genetics. The reassortant viruses replicated relatively poorly in embryonated chicken eggs. To improve virus growth in eggs, reassortants expressing the HA and NA of CA09 were passaged in MDCK cells and variants exhibiting large-plaque morphology were isolated. These variants replicated at levels approximately 10-fold higher than the rate of replication of the parental strains in embryonated chicken eggs. Sequence analysis indicated that single amino acid changes at positions 119, 153, 154, and 186 were responsible for the improved growth properties in MDCK cells and eggs. In addition, the introduction of a mutation at residue 155 that was previously shown to enhance the replication of a 1976 swine influenza virus also significantly improved the replication of the CA09 virus in eggs. Each variant was further evaluated for receptor binding preference, antigenicity, attenuation phenotype, and immunogenicity. Mutations at residues 153, 154, and 155 drastically reduced viral antigenicity, which made these mutants unsuitable as vaccine candidates. However, changes at residues 119 and 186 did not affect virus antigenicity or immunogenicity, justifying their inclusion in live attenuated vaccine candidates to protect against the currently circulating 2009 swine origin H1N1 viruses. PMID:19864389

  5. A Live Attenuated Equine H3N8 Influenza Vaccine Is Highly Immunogenic and Efficacious in Mice and Ferrets

    PubMed Central

    Baz, Mariana; Paskel, Myeisha; Matsuoka, Yumiko; Zengel, James; Cheng, Xing; Treanor, John J.; Jin, Hong

    2014-01-01

    ABSTRACT Equine influenza viruses (EIV) are responsible for rapidly spreading outbreaks of respiratory disease in horses. Although natural infections of humans with EIV have not been reported, experimental inoculation of humans with these viruses can lead to a productive infection and elicit a neutralizing antibody response. Moreover, EIV have crossed the species barrier to infect dogs, pigs, and camels and therefore may also pose a threat to humans. Based on serologic cross-reactivity of H3N8 EIV from different lineages and sublineages, A/equine/Georgia/1/1981 (eq/GA/81) was selected to produce a live attenuated candidate vaccine by reverse genetics with the hemagglutinin and neuraminidase genes of the eq/GA/81 wild-type (wt) virus and the six internal protein genes of the cold-adapted (ca) A/Ann Arbor/6/60 (H2N2) vaccine donor virus, which is the backbone of the licensed seasonal live attenuated influenza vaccine. In both mice and ferrets, intranasal administration of a single dose of the eq/GA/81 ca vaccine virus induced neutralizing antibodies and conferred complete protection from homologous wt virus challenge in the upper respiratory tract. One dose of the eq/GA/81 ca vaccine also induced neutralizing antibodies and conferred complete protection in mice and nearly complete protection in ferrets upon heterologous challenge with the H3N8 (eq/Newmarket/03) wt virus. These data support further evaluation of the eq/GA/81 ca vaccine in humans for use in the event of transmission of an equine H3N8 influenza virus to humans. IMPORTANCE Equine influenza viruses have crossed the species barrier to infect other mammals such as dogs, pigs, and camels and therefore may also pose a threat to humans. We believe that it is important to develop vaccines against equine influenza viruses in the event that an EIV evolves, adapts, and spreads in humans, causing disease. We generated a live attenuated H3N8 vaccine candidate and demonstrated that the vaccine was immunogenic and

  6. Technology for Independent Living: Sourcebook.

    ERIC Educational Resources Information Center

    Enders, Alexandra, Ed.

    This sourcebook provides information for the practical implementation of independent living technology in the everyday rehabilitation process. "Information Services and Resources" lists databases, clearinghouses, networks, research and development programs, toll-free telephone numbers, consumer protection caveats, selected publications, and…

  7. Living with Idiopathic Pulmonary Fibrosis

    MedlinePlus

    ... and Support Living with IPF may cause fear, anxiety, depression, and stress. Talk about how you feel ... and friends also can help relieve stress and anxiety. Let your loved ones know how you feel ...

  8. [Passive euthanasia and living will].

    PubMed

    Julesz, Máté

    2014-07-01

    This article deals with the intentional distinction between murder of first degree and passive euthanasia. In Hungary, active euthanasia is considered to be a murder of first degree, whilst the Netherlands, Belgium, Luxemburg and Switzerland have legalized the active form of mercy killing in Europe. The palliative terminal care, when e.g. giving pain-killer morphine to the patient, might result in decreasing the patient's life-span, and thus causing indirect euthanasia. However, the legal institution of living will exists in several counter-euthanasia countries. The living will allows future patients to express their decision in advance to refuse a life-sustaining treatment, e.g. in case of irreversible coma. The institution of living will exists in Germany and in Hungary too. Nevertheless, the formal criteria of living will make it hardly applicable. The patient ought to express his/her will before a notary public in advance, and he/she should hand it over when being hospitalized. If the patient is not able to present his/her living will to his/her doctor in the hospital, then his/her only hope remains that he/she has given a copy of the living will to the family doctor previously, and the family doctor will notify the hospital.

  9. Parallel molecular routes to cold adaptation in eight genera of New Zealand stick insects

    PubMed Central

    Dennis, Alice B.; Dunning, Luke T.; Sinclair, Brent J.; Buckley, Thomas R.

    2015-01-01

    The acquisition of physiological strategies to tolerate novel thermal conditions allows organisms to exploit new environments. As a result, thermal tolerance is a key determinant of the global distribution of biodiversity, yet the constraints on its evolution are not well understood. Here we investigate parallel evolution of cold tolerance in New Zealand stick insects, an endemic radiation containing three montane-occurring species. Using a phylogeny constructed from 274 orthologous genes, we show that stick insects have independently colonized montane environments at least twice. We compare supercooling point and survival of internal ice formation among ten species from eight genera, and identify both freeze tolerance and freeze avoidance in separate montane lineages. Freeze tolerance is also verified in both lowland and montane populations of a single, geographically widespread, species. Transcriptome sequencing following cold shock identifies a set of structural cuticular genes that are both differentially regulated and under positive sequence selection in each species. However, while cuticular proteins in general are associated with cold shock across the phylogeny, the specific genes at play differ among species. Thus, while processes related to cuticular structure are consistently associated with adaptation for cold, this may not be the consequence of shared ancestral genetic constraints. PMID:26355841

  10. Extreme preconditioning: cold adaptation through sea swimming as a means to improving surgical outcomes.

    PubMed

    Harper, C Mark

    2012-04-01

    The practice of sea bathing for its health benefits was popularised by Richard Russell in Regency Brighton during the 18th Century. Although the cures he claimed it could effect seem a little far-fetched today, as with many historical remedies, there is much to be gained from revisiting such theories in the light of modern medical research. In this paper I will draw parallels between the surgical stress response and the response to cold exposure and hypothesise how a programme of sea bathing may be used to enhance postoperative recovery and reduce preoperative complications.

  11. Comparative expression study to increase the solubility of cold adapted Vibrio proteins in Escherichia coli.

    PubMed

    Niiranen, Laila; Espelid, Sigrun; Karlsen, Christian R; Mustonen, Milla; Paulsen, Steinar M; Heikinheimo, Pirkko; Willassen, Nils P

    2007-03-01

    Functional and structural studies require gene overexpression and purification of soluble proteins. We wanted to express proteins from the psychrophilic bacterium Vibrio salmonicida in Escherichia coli, but encountered solubility problems. To improve the solubility of the proteins, we compared the effects of six N-terminal fusion proteins (Gb1, Z, thioredoxin, GST, MBP and NusA) and an N-terminal His6-tag. The selected test set included five proteins from the fish pathogen V. salmonicida and two related products from the mesophilic human pathogen Vibrio cholerae. We tested the expression in two different expression strains and at three different temperatures (16, 23 and 37 degrees C). His6-tag was the least effective tag, and these vector constructs were also difficult to transform. MBP and NusA performed best, expressing soluble proteins with all fusion partners in at least one of the cell types. In some cases MBP, GST and thioredoxin fusions resulted in products of incorrect size. The effect of temperature is complex: in most cases level of expression increased with temperature, whereas the effect on solubility was opposite. We found no clear connection between the preferred expression temperature of the protein and the temperature of the original host organism's natural habitat.

  12. The genome of the polar eukaryotic microalga Coccomyxa subellipsoidea reveals traits of cold adaptation

    SciTech Connect

    Blanc, Guillaume; Agarkova, Irina; Grimwood, Jane; Kuo, Alan; Brueggeman, Andrew; Dunigan, David D.; Gurnon, James; Ladunga, Istvan; Lindquist, Erika; Lucas, Susan; Pangilinan, Jasmyn; Proschold, Thomas; Salamov, Asaf; Schmutz, Jeremy; Weeks, Donald; Tamada, Takashi; Lomsadze, Alexandre; Borodovsky, Mark; Claverie, Jean-Michel; Grigoriev, Igor V.; Van Etten, James L.

    2012-02-13

    Background Little is known about the mechanisms of adaptation of life to the extreme environmental conditions encountered in polar regions. Here we present the genome sequence of a unicellular green alga from the division chlorophyta, Coccomyxa subellipsoidea C-169, which we will hereafter refer to as C-169. This is the first eukaryotic microorganism from a polar environment to have its genome sequenced. Results The 48.8 Mb genome contained in 20 chromosomes exhibits significant synteny conservation with the chromosomes of its relatives Chlorella variabilis and Chlamydomonas reinhardtii. The order of the genes is highly reshuffled within synteny blocks, suggesting that intra-chromosomal rearrangements were more prevalent than inter-chromosomal rearrangements. Remarkably, Zepp retrotransposons occur in clusters of nested elements with strictly one cluster per chromosome probably residing at the centromere. Several protein families overrepresented in C. subellipsoidae include proteins involved in lipid metabolism, transporters, cellulose synthases and short alcohol dehydrogenases. Conversely, C-169 lacks proteins that exist in all other sequenced chlorophytes, including components of the glycosyl phosphatidyl inositol anchoring system, pyruvate phosphate dikinase and the photosystem 1 reaction center subunit N (PsaN). Conclusions We suggest that some of these gene losses and gains could have contributed to adaptation to low temperatures. Comparison of these genomic features with the adaptive strategies of psychrophilic microbes suggests that prokaryotes and eukaryotes followed comparable evolutionary routes to adapt to cold environments.

  13. Pancreatic triacylglycerol lipase in a hibernating mammal. II. Cold-adapted function and differential expression.

    PubMed

    Squire, Teresa L; Lowe, Mark E; Bauer, Vernon W; Andrews, Matthew T

    2003-12-16

    Thirteen-lined ground squirrels (Spermophilus tridecemlineatus) exploit the low-temperature activity of pancreatic triacylglycerol lipase (PTL) during hibernation. Lipolytic activity at body temperatures associated with hibernation was examined using recombinant ground squirrel and human PTLs expressed in yeast. Both the human and ground squirrel enzymes displayed high activity at temperatures as low as 0 degrees C and showed Q10 values of 1.2-1.5 over a range of 37-7 degrees C. These studies indicate that low-temperature lipolysis is a general property of PTL and does not require protein modifications unique to mammalian cells and/or the hibernating state. Western blots show elevated levels of PTL protein during hibernation in both heart and white adipose tissue (WAT). Significant increases in PTL gene expression are seen in heart, WAT, and testes; but not in pancreas, where PTL mRNA levels are highest. Upregulation of PTL in testes is also accompanied by expression of the PTL-specific cofactor, colipase. The multi-tissue expression of PTL during hibernation supports its role as a key enzyme that shows high activity at low temperatures.

  14. Biodegradation of methyl tert-butyl ether by cold-adapted mixed and pure bacterial cultures.

    PubMed

    Zaitsev, G M; Uotila, J S; Häggblom, M M

    2007-04-01

    An aerobic mixed bacterial culture (CL-EMC-1) capable of utilizing methyl tert-butyl ether (MTBE) as the sole source of carbon and energy with a growth temperature range of 3 to 30 degrees C and optimum of 18 to 22 degrees C was enriched from activated sludge. Transient accumulation of tert-butanol (TBA) occurred during utilization of MTBE at temperatures from 3 degrees C to 14 degrees C, but TBA did not accumulate above 18 degrees C. The culture utilized MTBE at a concentration of up to 1.5 g l(-1) and TBA of up to 7 g l(-1). The culture grew on MTBE at a pH range of 5 to 9, with an optimum pH of 6.5 to 7.1. The specific growth rate of the CL-EMC-1 culture on 0.1 g l(-1) of MTBE at 22 degrees C and pH 7.1 was 0.012 h(-1), and the growth yield was 0.64 g (dry weight) g(-1). A new MTBE-utilizing bacterium, Variovorax paradoxus strain CL-8, isolated from the mixed culture utilized MTBE, TBA, 2-hydroxy isobutyrate, lactate, methacrylate, and acetate as sole sources of carbon and energy but not 2-propanol, acetone, methanol, formaldehyde, or formate. Two other isolates, Hyphomicrobium facilis strain CL-2 and Methylobacterium extorquens strain CL-4, isolated from the mixed culture were able to grow on C(1) compounds. The combined consortium could thus utilize all of the carbon of MTBE. PMID:17146651

  15. Protein surface softness is the origin of enzyme cold-adaptation of trypsin.

    PubMed

    Isaksen, Geir Villy; Åqvist, Johan; Brandsdal, Bjørn Olav

    2014-08-01

    Life has effectively colonized most of our planet and extremophilic organisms require specialized enzymes to survive under harsh conditions. Cold-loving organisms (psychrophiles) express heat-labile enzymes that possess a high specific activity and catalytic efficiency at low temperatures. A remarkable universal characteristic of cold-active enzymes is that they show a reduction both in activation enthalpy and entropy, compared to mesophilic orthologs, which makes their reaction rates less sensitive to falling temperature. Despite significant efforts since the early 1970s, the important question of the origin of this effect still largely remains unanswered. Here we use cold- and warm-active trypsins as model systems to investigate the temperature dependence of the reaction rates with extensive molecular dynamics free energy simulations. The calculations quantitatively reproduce the catalytic rates of the two enzymes and further yield high-precision Arrhenius plots, which show the characteristic trends in activation enthalpy and entropy. Detailed structural analysis indicates that the relationship between these parameters and the 3D structure is reflected by significantly different internal protein energy changes during the reaction. The origin of this effect is not localized to the active site, but is found in the outer regions of the protein, where the cold-active enzyme has a higher degree of softness. Several structural mechanisms for softening the protein surface are identified, together with key mutations responsible for this effect. Our simulations further show that single point-mutations can significantly affect the thermodynamic activation parameters, indicating how these can be optimized by evolution.

  16. Molecular dynamics of the salt dependence of a cold-adapted enzyme: endonuclease I.

    PubMed

    Benrezkallah, D; Dauchez, M; Krallafa, A M

    2015-01-01

    The effects of salt on the stability of globular proteins have been known for a long time. In the present investigations, we shall focus on the effect of the salt ions upon the structure and the activity of the endonuclease I enzyme. In the present work, we shall focus on the relationship between ion position and the structural features of the Vibrio salmonicida (VsEndA) enzyme. We will concentrate on major questions such as: how can salt ions affect the molecular structure? What is the activity of the enzyme and which specific regions are directly involved? For that purpose, we will study the behaviour of the VsEndA over different salt concentrations using molecular dynamics (MD) simulations. We report the results of MD simulations of the endonuclease I enzyme at five different salt concentrations. Analysis of trajectories in terms of the root mean square fluctuation (RMSF), radial distribution function, contact numbers and hydrogen bonding lifetimes, indicate distinct differences when changing the concentration of NaCl. Results are found to be in good agreement with experimental data, where we have noted an optimum salt concentration for activity equal to 425 mM. Under this salt concentration, the VsEndA exhibits two more flexible loop regions, compared to the other salt concentrations. When analysing the RMSF of these two specific regions, three residues were selected for their higher mobility. We find a correlation between the structural properties studied here such as the radial distribution function, the contact numbers and the hydrogen bonding lifetimes, and the structural flexibility of only two polar residues. Finally, in the light of the present work, the molecular basis of the salt adaptation of VsEndA enzyme has been explored by mean of explicit solvent and salt treatment. Our results reveal that modulation of the sodium/chloride ions interaction with some specific loop regions of the protein is the strategy followed by this type of psychrophilic enzyme to enhance catalytic activity at the physiological conditions.

  17. Legionella species diversity and dynamics from surface reservoir to tap water: from cold adaptation to thermophily.

    PubMed

    Lesnik, René; Brettar, Ingrid; Höfle, Manfred G

    2016-05-01

    Water samples of the Drinking Water Supply System (DWSS) of the city of Braunschweig were analysed for its Legionella species composition using genus-specific PCR amplicons and single-strand conformation polymorphism (SSCP) fingerprint analyses based on 16S rRNA genes. These analyses comprised the whole supply chain including raw water, treatment process and large-scale storage, and a seasonal study of finished drinking water sampled monthly from cold and hot tap water. Treatment of raw water had a major impact on Legionella species by reducing their diversity and abundances. The Legionella species composition of the tap water was highly distinct from that of both source waters. In cold water, 8-14 different phylotypes of Legionella (PTLs) were observed per sample with relative abundances ranging from >1% to 53%. In hot water, L. pneumophila was present during all seasons at high relative abundances (8-40%) accompanied by 5-14 other PTLs of which 6 PTLs were in common with cold water. This thermophilic Legionella community, including L. pneumophila, was able to grow in the hot water above 50 °C. Such thermophilic Legionella populations are of general relevance for drinking water management and public health, but also for the ecology and evolution of the genus Legionella. PMID:26528838

  18. Protein surface softness is the origin of enzyme cold-adaptation of trypsin.

    PubMed

    Isaksen, Geir Villy; Åqvist, Johan; Brandsdal, Bjørn Olav

    2014-08-01

    Life has effectively colonized most of our planet and extremophilic organisms require specialized enzymes to survive under harsh conditions. Cold-loving organisms (psychrophiles) express heat-labile enzymes that possess a high specific activity and catalytic efficiency at low temperatures. A remarkable universal characteristic of cold-active enzymes is that they show a reduction both in activation enthalpy and entropy, compared to mesophilic orthologs, which makes their reaction rates less sensitive to falling temperature. Despite significant efforts since the early 1970s, the important question of the origin of this effect still largely remains unanswered. Here we use cold- and warm-active trypsins as model systems to investigate the temperature dependence of the reaction rates with extensive molecular dynamics free energy simulations. The calculations quantitatively reproduce the catalytic rates of the two enzymes and further yield high-precision Arrhenius plots, which show the characteristic trends in activation enthalpy and entropy. Detailed structural analysis indicates that the relationship between these parameters and the 3D structure is reflected by significantly different internal protein energy changes during the reaction. The origin of this effect is not localized to the active site, but is found in the outer regions of the protein, where the cold-active enzyme has a higher degree of softness. Several structural mechanisms for softening the protein surface are identified, together with key mutations responsible for this effect. Our simulations further show that single point-mutations can significantly affect the thermodynamic activation parameters, indicating how these can be optimized by evolution. PMID:25165981

  19. Parallel molecular routes to cold adaptation in eight genera of New Zealand stick insects.

    PubMed

    Dennis, Alice B; Dunning, Luke T; Sinclair, Brent J; Buckley, Thomas R

    2015-09-10

    The acquisition of physiological strategies to tolerate novel thermal conditions allows organisms to exploit new environments. As a result, thermal tolerance is a key determinant of the global distribution of biodiversity, yet the constraints on its evolution are not well understood. Here we investigate parallel evolution of cold tolerance in New Zealand stick insects, an endemic radiation containing three montane-occurring species. Using a phylogeny constructed from 274 orthologous genes, we show that stick insects have independently colonized montane environments at least twice. We compare supercooling point and survival of internal ice formation among ten species from eight genera, and identify both freeze tolerance and freeze avoidance in separate montane lineages. Freeze tolerance is also verified in both lowland and montane populations of a single, geographically widespread, species. Transcriptome sequencing following cold shock identifies a set of structural cuticular genes that are both differentially regulated and under positive sequence selection in each species. However, while cuticular proteins in general are associated with cold shock across the phylogeny, the specific genes at play differ among species. Thus, while processes related to cuticular structure are consistently associated with adaptation for cold, this may not be the consequence of shared ancestral genetic constraints.

  20. Dynamics of some parameters of the endocrine and lymphatic systems in rats during cold adaptation

    SciTech Connect

    Borodin, Yu.I.; Sedova, L.A.; Selyatitskaya, V.G.; Shorin, Yu.P.

    1986-02-01

    This paper examines the combined behavior of the endocrine and lymphatic systems in rats at stages of long-term adaptation of the animals to moderate cold. After decapitation of male Wister rats, the corticosterone concentration in the blood plasma was determined by saturation analysis and serum levels of thyroxine (T/sub 4/) and triiodothyronine (T/sub 3/) were determined by radioimmunoassay. The thymus was weighed and the structure of the popliteal lymph nodes (LN) was studied in histological sections stained with hematoxylin and eosin and with azure II-eosin. Morphometry of the structural components of LN was undertaken and the numbers of the various cell forms per 1000 cells were counted in different zones of LN. The increase in activity of the lymphoid tissue in the phase of adaptation may be connected with intensification of the peripheral action of thyroid hormones. During long-term adaptation, in the phase of consistently increased specific resistance, a new type of endocrine-lymphoid relation is formed, and it differs significantly both in the original state and in the acute phase of stress.

  1. Video fingerprinting for live events

    NASA Astrophysics Data System (ADS)

    Celik, Mehmet; Haitsma, Jaap; Barvinko, Pavlo; Langelaar, Gerhard; Maas, Martijn

    2009-02-01

    Multimedia fingerprinting (robust hashing) as a content identification technology is emerging as an effective tool for preventing unauthorized distribution of commercial content through user generated content (UGC) sites. Research in the field has mainly considered content types with slow distribution cycles, e.g. feature films, for which reference fingerprint ingestion and database indexing can be performed offline. As a result, research focus has been on improving the robustness and search speed. Live events, such as live sports broadcasts, impose new challenges on a fingerprinting system. For instance, highlights from a soccer match are often available-and viewed-on UGC sites well before the end of the match. In this scenario, the fingerprinting system should be able to ingest and index live content online and offer continuous search capability, where new material is identifiable within minutes of broadcast. In this paper, we concentrate on algorithmic and architectural challenges we faced when developing a video fingerprinting solution for live events. In particular, we discuss how to effectively utilize fast sorting algorithms and a master-slave architecture for fast and continuous ingestion of live broadcasts.

  2. The Matrix Gene Segment Destabilizes the Acid and Thermal Stability of the Hemagglutinin of Pandemic Live Attenuated Influenza Virus Vaccines

    PubMed Central

    O'Donnell, Christopher D.; Vogel, Leatrice; Matsuoka, Yumiko; Jin, Hong

    2014-01-01

    ABSTRACT The threat of future influenza pandemics and their potential for rapid spread, morbidity, and mortality has led to the development of pandemic vaccines. We generated seven reassortant pandemic live attenuated influenza vaccines (pLAIVs) with the hemagglutinin (HA) and neuraminidase (NA) genes derived from animal influenza viruses on the backbone of the six internal protein gene segments of the temperature sensitive, cold-adapted (ca) A/Ann Arbor/60 (H2N2) virus (AA/60 ca) of the licensed seasonal LAIV. The pLAIV viruses were moderately to highly restricted in replication in seronegative adults; we sought to determine the biological basis for this restriction. Avian influenza viruses generally replicate at higher temperatures than human influenza viruses and, although they shared the same backbone, the pLAIV viruses had a lower shutoff temperature than seasonal LAIV viruses, suggesting that the HA and NA influence the degree of temperature sensitivity. The pH of HA activation of highly pathogenic avian influenza viruses was greater than human and low-pathogenicity avian influenza viruses, as reported by others. However, pLAIV viruses had a consistently higher pH of HA activation and reduced HA thermostability compared to the corresponding wild-type parental viruses. From studies with single-gene reassortant viruses bearing one gene segment from the AA/60 ca virus in recombinant H5N1 or pH1N1 viruses, we found that the lower HA thermal stability and increased pH of HA activation were associated with the AA/60 M gene. Together, the impaired HA acid and thermal stability and temperature sensitivity likely contributed to the restricted replication of the pLAIV viruses we observed in seronegative adults. IMPORTANCE There is increasing evidence that the HA stability of influenza viruses depends on the virus strain and host species and that HA stability can influence replication, virulence, and transmission of influenza A viruses in different species. We

  3. Information use in colonial living.

    PubMed

    Evans, Julian C; Votier, Stephen C; Dall, Sasha R X

    2016-08-01

    Despite the fact that many animals live in groups, there is still no clear consensus about the ecological or evolutionary mechanisms underlying colonial living. Recently, research has suggested that colonies may be important as sources of social information. The ready availability of information from conspecifics allows animals to make better decisions about avoiding predators, reducing brood parasitism, migratory phenology, mate choice, habitat choice and foraging. These choices can play a large part in the development and maintenance of colonies. Here we review the types of information provided by colonial animals and examine the different ways in which decision-making in colonies can be enhanced by social information. We discuss what roles information might take in the evolution, formation and maintenance of colonies. In the process, we illustrate that information use permeates all aspects of colonial living.

  4. Live Imaging of the Lung

    PubMed Central

    Looney, Mark R.; Bhattacharya, Jahar

    2015-01-01

    Live lung imaging has spanned the discovery of capillaries in the frog lung by Malpighi to the current use of single and multiphoton imaging of intravital and isolated perfused lung preparations incorporating fluorescent molecular probes and transgenic reporter mice. Along the way, much has been learned about the unique microcirculation of the lung, including immune cell migration and the mechanisms by which cells at the alveolar-capillary interface communicate with each other. In this review, we highlight live lung imaging techniques as applied to the role of mitochondria in lung immunity, mechanisms of signal transduction in lung compartments, studies on the composition of alveolar wall liquid, and neutrophil and platelet trafficking in the lung under homeostatic and inflammatory conditions. New applications of live lung imaging and the limitations of current techniques are discussed. PMID:24245941

  5. How to increase living donation.

    PubMed

    Davis, Connie L

    2011-04-01

    Living donation is the key to increasing access to successful solid organ transplantation worldwide. However, the means to expanding the number of living donors on a global scale are not known. Although there have been many suggestions for the best approach, cultural issues may limit the effectiveness of some strategies. Only a few ideas have been studied, and one in particular- outright payment to donors - may raise ethical issues that are difficult to surmount and might negatively alter altruistic behavior. With respect to the present environment, this article will describe some of the approaches that are being discussed to increase the number of living donors, with a particular focus on kidney transplantation. PMID:21210867

  6. Microencapsulation of live probiotic bacteria.

    PubMed

    Islam, Mohammad Ariful; Yun, Cheol-Heui; Choi, Yun-Jaie; Cho, Chong-Su

    2010-10-01

    Scientific research regarding the use of live bacterial cells for therapeutic purposes has been rapidly growing over the years and has generated considerable interest to scientists and health professionals. Probiotics are defined as essential live microorganisms which, when administered in adequate amounts, confer a health benefit on the host. Due to considerable beneficial health effects, these microorganisms are increasingly incorporated into the dairy products; however, many reports demonstrated their poor survival and stability. Their survival in the gastrointestinal (GI) tract is also questionable. To overcome these problems, microencapsulation techniques are currently receiving considerable attention. This review describes the importance of live probiotic bacterial microencapsulation using an alginate microparticulate system and presents the potentiality of various coating polymers such as chitosan and polylysine for improving the stability of this microencapsulation. PMID:21030820

  7. [Health effects of living habits].

    PubMed

    Vuori, Ilkka

    2015-01-01

    Single healthy living habits such as non-smoking and regular physical activity decrease the risk of common non-communicable diseases, unsuccessful aging and premature death to a small to moderate degree. Their cumulative effects are, however, large. Only a small minority of people adhere well to all healthy living habits or even the healthiest ones. Consequently, the population attributable fractions of major public health problems due to unhealthy lifestyles are large. Substantial improvement of public health calls for policies and programs to influence the root causes of the lifestyles in the multiple environments and systems where they are developed, maintained, and changed.

  8. Astronomy Cast Live: Live Blogging Today's Science to the World

    NASA Astrophysics Data System (ADS)

    Bemrose-Fetter, Rebecca; Gay, P. L.; Astronomy Cast LIVE Team

    2008-05-01

    In today's digital, on-demand society, consumers of information both want to know exactly what is happening as it is happening, and to be able to subscribe to content of their choosing. Meeting the needs of these tech savvy individuals are bloggers, podcasters and vodcasters. Using text, audio, and video to reach their respective audiences, these communicators are the new face of public outreach and journalism, but even their communications means are starting to become passé in the face of live blogging. The idea behind live blogging is simple: Take any person - even an undergraduate - with an Internet connected device, put them someplace interesting, and have them report on what they are seeing and experiencing online in real-time. This new tool is bringing astronomy enthusiasts around the world the thrill of live astronomy announcements, attending talks in real, and being "in the room" with astronomers via an Internet connection. These audiences can be anyone, from any nation, with any age. Beyond the public communications benefits of this program, it is also a program that allows the participation of early undergraduate students in science conferences. To date, two undergraduate students and five E/PO professionals have live blogged text, audio, and video content from three science conferences and a shuttle launch. Together, they have produced over 200 hundred stories that have reached tens of thousands of people around the world. In this poster we describe how we have made astronomy live blogging a reality from both the technical and personal standpoint. This project is funded through NSF grant # 0744944.

  9. A live attenuated H7N7 candidate vaccine virus induces neutralizing antibody that confers protection from challenge in mice, ferrets, and monkeys.

    PubMed

    Min, Ji-Young; Vogel, Leatrice; Matsuoka, Yumiko; Lu, Bin; Swayne, David; Jin, Hong; Kemble, George; Subbarao, Kanta

    2010-11-01

    A live attenuated H7N7 candidate vaccine virus was generated by reverse genetics using the modified hemagglutinin (HA) and neuraminidase (NA) genes of highly pathogenic (HP) A/Netherlands/219/03 (NL/03) (H7N7) wild-type (wt) virus and the six internal protein genes of the cold-adapted (ca) A/Ann Arbor/6/60 ca (AA ca) (H2N2) virus. The reassortant H7N7 NL/03 ca vaccine virus was temperature sensitive and attenuated in mice, ferrets, and African green monkeys (AGMs). Intranasal (i.n.) administration of a single dose of the H7N7 NL/03 ca vaccine virus fully protected mice from lethal challenge with homologous and heterologous H7 viruses from Eurasian and North American lineages. Two doses of the H7N7 NL/03 ca vaccine induced neutralizing antibodies in serum and provided complete protection from pulmonary replication of homologous and heterologous wild-type H7 challenge viruses in mice and ferrets. One dose of the H7N7 NL/03 ca vaccine elicited an antibody response in one of three AGMs that was completely protected from pulmonary replication of the homologous wild-type H7 challenge virus. The contribution of CD8(+) and/or CD4(+) T cells to the vaccine-induced protection of mice was evaluated by T-cell depletion; T lymphocytes were not essential for the vaccine-induced protection from lethal challenge with H7 wt viruses. Additionally, passively transferred neutralizing antibody induced by the H7N7 NL/03 ca virus protected mice from lethality following challenge with H7 wt viruses. The safety, immunogenicity, and efficacy of the H7N7 NL/03 ca vaccine virus in mice, ferrets, and AGMs support the evaluation of this vaccine virus in phase I clinical trials. PMID:20810733

  10. Live Attenuated Influenza Vaccine Strains Elicit a Greater Innate Immune Response than Antigenically-Matched Seasonal Influenza Viruses during Infection of Human Nasal Epithelial Cell Cultures

    PubMed Central

    Fischer, William A.; Brighton, Missy; Jaspers, Ilona

    2014-01-01

    Influenza viruses are global pathogens that infect approximately 10–20% of the world’s population each year. Vaccines, including the live attenuated influenza vaccine (LAIV), are the best defense against influenza infections. The LAIV is a novel vaccine that actively replicates in the human nasal epithelium and elicits both mucosal and systemic protective immune responses. The differences in replication and innate immune responses following infection of human nasal epithelium with influenza seasonal wild type (WT) and LAIV viruses remain unknown. Using a model of primary differentiated human nasal epithelial cell (hNECs) cultures, we compared influenza WT and antigenically-matched cold adapted (CA) LAIV virus replication and the subsequent innate immune response including host cellular pattern recognition protein expression, host innate immune gene expression, secreted pro-inflammatory cytokine production, and intracellular viral RNA levels. Growth curves comparing virus replication between WT and LAIV strains revealed significantly less infectious virus production during LAIV compared with WT infection. Despite this disparity in infectious virus production the LAIV strains elicited a more robust innate immune response with increased expression of RIG-I, TLR-3, IFNβ, STAT-1, IRF-7, MxA, and IP-10. There were no differences in cytotoxicity between hNEC cultures infected with WT and LAIV strains as measured by basolateral levels of LDH. Elevated levels of intracellular viral RNA during LAIV as compared with WT virus infection of hNEC cultures at 33°C may explain the augmented innate immune response via the up-regulation of pattern recognition receptors and down-stream type I IFN expression. Taken together our results suggest that the decreased replication of LAIV strains in human nasal epithelial cells is associated with a robust innate immune response that differs from infection with seasonal influenza viruses, limits LAIV shedding and plays a role in the

  11. Live attenuated influenza vaccine strains elicit a greater innate immune response than antigenically-matched seasonal influenza viruses during infection of human nasal epithelial cell cultures.

    PubMed

    Fischer, William A; Chason, Kelly D; Brighton, Missy; Jaspers, Ilona

    2014-03-26

    Influenza viruses are global pathogens that infect approximately 10-20% of the world's population each year. Vaccines, including the live attenuated influenza vaccine (LAIV), are the best defense against influenza infections. The LAIV is a novel vaccine that actively replicates in the human nasal epithelium and elicits both mucosal and systemic protective immune responses. The differences in replication and innate immune responses following infection of human nasal epithelium with influenza seasonal wild type (WT) and LAIV viruses remain unknown. Using a model of primary differentiated human nasal epithelial cell (hNECs) cultures, we compared influenza WT and antigenically-matched cold adapted (CA) LAIV virus replication and the subsequent innate immune response including host cellular pattern recognition protein expression, host innate immune gene expression, secreted pro-inflammatory cytokine production, and intracellular viral RNA levels. Growth curves comparing virus replication between WT and LAIV strains revealed significantly less infectious virus production during LAIV compared with WT infection. Despite this disparity in infectious virus production the LAIV strains elicited a more robust innate immune response with increased expression of RIG-I, TLR-3, IFNβ, STAT-1, IRF-7, MxA, and IP-10. There were no differences in cytotoxicity between hNEC cultures infected with WT and LAIV strains as measured by basolateral levels of LDH. Elevated levels of intracellular viral RNA during LAIV as compared with WT virus infection of hNEC cultures at 33°C may explain the augmented innate immune response via the up-regulation of pattern recognition receptors and down-stream type I IFN expression. Taken together our results suggest that the decreased replication of LAIV strains in human nasal epithelial cells is associated with a robust innate immune response that differs from infection with seasonal influenza viruses, limits LAIV shedding and plays a role in the silent

  12. Project in Apartment Group Living

    ERIC Educational Resources Information Center

    Riehman, Lynne; O'Brien, Carolyn F.

    1973-01-01

    After discharge from the hospital, many mental patients go back to the kind of environment that contributed to their breakdown. In the belief that new options should be available to mental patients when discharged, a hospital in New York City developed--with marked success--a plan for group living. (Author)

  13. Living Assessment Passes the Test

    ERIC Educational Resources Information Center

    Suskind, Dorothy C.

    2015-01-01

    The author, a 5th-grade teacher at an independent boys' school, gives a first-person account of how her constant assessments and requirement that her students be active participants in their own learning gainsays the need for high-stakes, standardized testing. She posits a "living assessment" that is intertwined, interactive and…

  14. Living History: F. Eugene Yates

    ERIC Educational Resources Information Center

    Urquhart, John

    2009-01-01

    In 2005, the American Physiological Society (APS) initiated the Living History of Physiology Archival Program to recognize senior members who have made significant contributions during their career to the advancement of the discipline and the profession of physiology. During 2008, the APS Cardiovascular Section selected Francis Eugene Yates to be…

  15. Senior to Senior: Living Lessons

    ERIC Educational Resources Information Center

    Goff, Kathy

    2004-01-01

    Senior to Senior: Living Lessons is a program created to provide meaningful horticulture therapy activities for community minority elders (60 years of age and older) and senior college students (20 years of age and older) from an Historically Black University. The program's objectives were to promote positive intergenerational relationships and to…

  16. College for Living Instructor's Manual.

    ERIC Educational Resources Information Center

    Templin, Robert G., Jr.; And Others

    This five-part manual was designed to help volunteer instructors in Northern Virginia Community College's College for Living Program to conduct survival and socialization courses for handicapped adults. After introductory material summarizing general principles and specific suggestions, Robert Templin provides information on the skills and…

  17. Living Arrangements in Later Life.

    ERIC Educational Resources Information Center

    Frazier, Billie H.

    This document contains a brief bibliography of peer-reviewed literature, with abstracts, on living arrangements in later life. It is one of 12 bibliographies on aging prepared by the National Agricultural Library for its "Pathfinders" series of publications. Topics covered by the other 11 bibliographies include aging parents, adult children,…

  18. Shakespeare Live! and Character Counts.

    ERIC Educational Resources Information Center

    Brookshire, Cathy A.

    This paper discusses a live production of Shakespeare's "Macbeth" (in full costume but with no sets) for all public middle school and high school students in Harrisonburg and Rockingham, Virginia. The paper states that the "Character Counts" issues that are covered in the play are: decision making, responsibility and citizenship, trustworthiness,…

  19. Living History: Elsworth R. Buskirk

    ERIC Educational Resources Information Center

    Tipton, Charles M.

    2009-01-01

    In 2005, the American Physiological Society (APS) initiated the Living History of Physiology Archival Program to recognize senior members who have made significant contributions during their career to the advancement of the discipline and the profession of physiology. Subsequently, the leadership of the APS Section of Environmental and Exercise…

  20. Investigating Evolution with Living Plants.

    ERIC Educational Resources Information Center

    Schlessman, Mark A.

    1997-01-01

    Describes two investigative labs that use live plants to illustrate important biological principles, include quantitative analysis, and require very little equipment. Each lab is adaptable to a variety of class sizes, course contents, and student backgrounds. Topics include the evolution of flower size in Mimulus and pollination of Brassicas. (DDR)

  1. Chicanas: Their Voices, Their Lives.

    ERIC Educational Resources Information Center

    Lockert, Lucia Fox, Ed.

    This book is an oral history of ten Chicana women ranging in ages from 84 to 24. The collection of interviews reflects how their hard work and determination have significantly changed their lives for the better. The backgrounds of the women vary; some were born in Mexico and moved to the United States. Others were born in the Southwest and later…

  2. Assisted Living Services and Amenities

    MedlinePlus

    ... good Alzheimer’s care. Safety and Security Peace of mind drives many decisions to seek senior housing for yourself or a loved one. Having a secure building where you know you or your loved one is protected from wandering or emergencies is very important. Senior living residences ...

  3. Supramolecular polymerization: Living it up

    NASA Astrophysics Data System (ADS)

    Würthner, Frank

    2014-03-01

    Protein fibril formation is involved in many human diseases and thus has been mechanistically elucidated in the context of understanding -- and in turn treating -- them. This biological phenomenon has now also inspired the design of a supramolecular system that undergoes living polymerization.

  4. Finding a Place to Live.

    ERIC Educational Resources Information Center

    NatureScope, 1985

    1985-01-01

    Provides background information and student activities on bird habitats, how birds have adapted to living in these habitats, and bird migration. Each activity includes an objective, recommended age level(s), subject area(s), list of materials needed, and procedures. Ready-to-copy student materials (puzzles and worksheets) are included. (JN)

  5. I Know Where They Live.

    ERIC Educational Resources Information Center

    Worsley, Ed, Ed.

    This science-oriented reading readiness text is provided in two versions: English and Navajo. It consists of large black-and-white drawings of animals, birds, and insects familiar to the Southwestern U.S. One picture is provided on each page, labeled with the following simple sentences: "This is a [name of the animal]. He lives in [name of…

  6. Living with High Blood Pressure

    MedlinePlus

    ... page from the NHLBI on Twitter. Living With High Blood Pressure If you have high blood pressure, the best thing to do is to talk ... help you track your blood pressure. Pregnancy Planning High blood pressure can cause problems for mother and baby. High ...

  7. Living kidney donors and ESRD.

    PubMed

    Ross, Lainie Friedman

    2015-07-01

    There are more than 325 living kidney donors who have developed end-stage renal disease and have been listed on the Organ Procurement and Transplantation Network (OPTN)/United Network for Organ Sharing (UNOS) deceased donor kidney wait list. The OPTN/UNOS database records where these kidney donors are listed and, if they donated after April 1994, where that donation occurred. These 2 locations are often not the same. In this commentary, I examine whether a national living donor registry should be created and whether transplantation centers should be notified when one of their living kidney donors develops end-stage renal disease. I consider and refute 5 potential objections to center notification. I explain that transplantation centers should look back at these cases and input data into a registry to attempt to identify patterns that could improve donor evaluation protocols. Creating a registry and mining the information it contains is, in my view, our moral and professional responsibility to future patients and the transplantation endeavor. As individuals and as a community, we need to acknowledge the many unknown risks of living kidney donation and take responsibility for identifying these risks. We then must share information about these risks, educate prospective donors about them, and attempt to minimize them.

  8. Living in the Gifted Family

    ERIC Educational Resources Information Center

    DeVries, Arlene R.

    2010-01-01

    Raising gifted children is both a joy and a challenge. How does one survive and thrive living in a gifted family? Parents play an essential role in helping children develop appreciation and respect for the world and their place in it. Intellectual development and emotional reactions begin at an early age, and many major behavioral patterns are set…

  9. Living History: Clark M. Blatteis

    ERIC Educational Resources Information Center

    Quan, Ning

    2009-01-01

    In 2005, the American Physiological Society (APS) initiated the Living History Project to recognize senior members who have made extraordinary contributions during their career to the advancement of the discipline and profession of physiology. During 2007, the APS Section of Environmental and Exercise Physiology selected Clark M. Blatteis to be…

  10. Educating Lives for Christian Wisdom

    ERIC Educational Resources Information Center

    Davis, Darin H.; Wadell, Paul J.

    2016-01-01

    This article explores how educating lives for Christian wisdom might serve as an antidote to the vice of "acedia," a prominent feature of the culture of contemporary higher education. After suggesting that the capital vice of "acedia" seems to capture well various facets of our present age and how the pursuit of wisdom serves…

  11. Improvement of influenza A/Fujian/411/02 (H3N2) virus growth in embryonated chicken eggs by balancing the hemagglutinin and neuraminidase activities, using reverse genetics.

    PubMed

    Lu, Bin; Zhou, Helen; Ye, Dan; Kemble, George; Jin, Hong

    2005-06-01

    The H3N2 influenza A/Fujian/411/02-like virus strains that circulated during the 2003-2004 influenza season caused influenza epidemics. Most of the A/Fujian/411/02 virus lineages did not replicate well in embryonated chicken eggs and had to be isolated originally by cell culture. The molecular basis for the poor replication of A/Fujian/411/02 virus was examined in this study by the reverse genetics technology. Two antigenically related strains that replicated well in embryonated chicken eggs, A/Sendai-H/F4962/02 and A/Wyoming/03/03, were compared with the prototype A/Fujian/411/02 virus. A/Sendai differed from A/Fujian by three amino acids in the neuraminidase (NA), whereas A/Wyoming differed from A/Fujian by five amino acids in the hemagglutinin (HA). The HA and NA segments of these three viruses were reassorted with cold-adapted A/Ann Arbor/6/60, the master donor virus for the live attenuated type A influenza vaccines (FluMist). The HA and NA residues differed between these three H3N2 viruses evaluated for their impact on virus replication in MDCK cells and in embryonated chicken eggs. It was determined that replication of A/Fujian/411/02 in eggs could be improved by either changing minimum of two HA residues (G186V and V226I) to increase the HA receptor-binding ability or by changing a minimum of two NA residues (E119Q and Q136K) to lower the NA enzymatic activity. Alternatively, recombinant A/Fujian/411/02 virus could be adapted to grow in eggs by two amino acid substitutions in the HA molecule (H183L and V226A), which also resulted in the increased HA receptor-binding activity. Thus, the balance between the HA and NA activities is critical for influenza virus replication in a different host system. The HA or NA changes that increased A/Fujian/411/02 virus replication in embryonated chicken eggs were found to have no significant impact on antigenicity of these recombinant viruses. This study demonstrated that the reverse genetics technology could be used to

  12. Analyzing lysosomes in live cells.

    PubMed

    Pryor, Paul R

    2012-01-01

    Lysosomes are an important cellular organelle that receive and degrade macromolecules from the secretory, endocytic, autophagic, and phagocytic membrane-trafficking pathways. Defects in lysosome function lead to the development of disease with often-severe consequences to the individual. Since the discovery of lysosomes by Christian de Duve over 50 years ago, research into endocytic and lysosomal biology has allowed for the development of tools to understand further the role of lysosomes in cells. There are now several fluorescent probes that can be used to visualize and assess membrane traffic to the lysosome as well as probes to assess the activity of lysosomal hydrolases in live cells. This chapter describes the current methods used to measure lysosome function in live cells.

  13. [Living donor transplantation. Surgical complications].

    PubMed

    Karam, Georges

    2008-02-01

    Although nephrectomy by open surgery is the most used technique for the extraction of kidney transplants in the living donor, nephrectomy under laparaoscopy is increasingly practiced. Laparoscopic nephrectomy is less invasive and performed under videoscopy control, after insufflation of the peritoneal cavity. Three to four incisions are done in order to enter the surgical instruments. The kidney is extracted through a horizontal sus-pubic incision. The exposition is either exclusively transperitoneal, retroperitoneal or hand assisted. The advantages of laparoscopy are esthetical, financial due to a shorter hospitalisation and a quicker recovery, as well a confort for the donor. The disadvantages are a longer warm ischemia time and possibly a higher risk of delayed graft function. Randomised studies having compared laparoscopy and open surgery in the living donor have not find any significant difference regarding the per- and perioperative in the complications.

  14. [Living donor transplantation. Surgical complications].

    PubMed

    Karam, Georges

    2008-02-01

    Although nephrectomy by open surgery is the most used technique for the extraction of kidney transplants in the living donor, nephrectomy under laparaoscopy is increasingly practiced. Laparoscopic nephrectomy is less invasive and performed under videoscopy control, after insufflation of the peritoneal cavity. Three to four incisions are done in order to enter the surgical instruments. The kidney is extracted through a horizontal sus-pubic incision. The exposition is either exclusively transperitoneal, retroperitoneal or hand assisted. The advantages of laparoscopy are esthetical, financial due to a shorter hospitalisation and a quicker recovery, as well a confort for the donor. The disadvantages are a longer warm ischemia time and possibly a higher risk of delayed graft function. Randomised studies having compared laparoscopy and open surgery in the living donor have not find any significant difference regarding the per- and perioperative in the complications. PMID:18160357

  15. Transcription Dynamics in Living Cells.

    PubMed

    Lenstra, Tineke L; Rodriguez, Joseph; Chen, Huimin; Larson, Daniel R

    2016-07-01

    The transcription cycle can be roughly divided into three stages: initiation, elongation, and termination. Understanding the molecular events that regulate all these stages requires a dynamic view of the underlying processes. The development of techniques to visualize and quantify transcription in single living cells has been essential in revealing the transcription kinetics. They have revealed that (a) transcription is heterogeneous between cells and (b) transcription can be discontinuous within a cell. In this review, we discuss the progress in our quantitative understanding of transcription dynamics in living cells, focusing on all parts of the transcription cycle. We present the techniques allowing for single-cell transcription measurements, review evidence from different organisms, and discuss how these experiments have broadened our mechanistic understanding of transcription regulation.

  16. A Heart Set on Living.

    PubMed

    Berlin, Ana

    2016-01-01

    Quality of life is a highly subjective element on which to base health care decision making. This narrative reflection after the death of a family member uses poetry as a prompt to explore themes related to quality of life-including symptom burden, interpersonal relationships in the face of illness, and the will to live. Through penetrating inquiry and reflection, physicians and other care providers can gain insight into the underlying motivations, loyalties, and abilities that lend meaning to patients' lives and shape attitudes toward death and dying. By better recognizing and appreciating these factors, clinicians can develop patient-centered quality-of-life constructs that empower them to honor patient goals and preferences at the end of life. Physicians are encouraged to explore poetry and other artistic media to help foster the reflective capacity required to deeply understand and faithfully serve patients in this regard.

  17. Focusing light through living tissue

    NASA Astrophysics Data System (ADS)

    Vellekoop, I. M.; Aegerter, C. M.

    2010-02-01

    Tissues such as skin, fat or cuticle are non-transparent because inhomogeneities in the tissue scatter light. We demonstrate experimentally that light can be focused through turbid layers of living tissue, in spite of scattering. Our method is based on the fact that coherent light forms an interference pattern, even after hundreds of scattering events. By spatially shaping the wavefront of the incident laser beam, this interference pattern was modified to make the scattered light converge to a focus. In contrast to earlier experiments, where light was focused through solid objects, we focused light through living pupae of Drosophila melanogaster. We discuss a dynamic wavefront shaping algorithm that follows changes due to microscopic movements of scattering particles in real time. We relate the performance of the algorithm to the measured timescale of the changes in the speckle pattern and analyze our experiment in the light of Laser Doppler flowmetry. Applications in particle tracking, imaging, and optical manipulation are discussed.

  18. Steps to Independent Living Series.

    ERIC Educational Resources Information Center

    Lobb, Nancy

    This set of six activity books and a teacher's guide is designed to help students from eighth grade to adulthood with special needs to learn independent living skills. The activity books have a reading level of 2.5 and address: (1) "How to Get Well When You're Sick or Hurt," including how to take a temperature, see a doctor, and use medicines…

  19. Viruses: are they living entities?

    PubMed

    Pennazio, Sergio

    2011-01-01

    The essence (living or nonliving entities) of viruses has today become an aporia, i.e. a difficulty inherent in reasoning because they shared four fundamental characteristics with livings (multiplication, genetic information, mutation and evolution) without having the capacity to have an independent life. For much time, however, they were considered minuscule pathogenetic micro-organisms in observance of Koch and Pasteur's 'germ theory' albeit no microbiologist could show their existence except their filterability and pathogenetic action. Only some voices based on experimental results raised against this dogmatic view, in particular those of Beijerinck, Baur and Mrowka, without dipping effectively into the dominant theory. The discovery relative to their nucleoprotein nature made between 1934 and 1936 (Schlesinger as for the phage, and Bawden and co-operators as for Tobacco mosaic virus; TMV), together with the first demonstrations of their structures thanks to electron microscopy (from 1939 onwards) started on casting a new light on their true identity, which could be more clearly identified when, from 1955 onwards, phage and TMV proved to be decisive factors to understand the strategies of replication of the genetic material. Following the new knowledge, the theoretical view relative to viruses changed rather radically and the current view looks on these pathogenetic agents as nonliving aggregates of macromolecules provided with biological properties. There is, however, a current of thought, made explicitly by Lwoff that places viruses as compromise between living and non living and, perhaps, as primitive forms of life which have had great importance for the evolution of cellular life. At any rate, viruses are peculiar entities whose importance cannot be unacknowledged. PMID:22220354

  20. Exact law of live nature

    NASA Astrophysics Data System (ADS)

    Azbel, Mark Ya.

    2005-07-01

    Exact law of mortality dynamics in changing populations and environment is derived. It includes no explicit characteristics of animal- environment interactions (metabolism etc) which are a must for life; it is universal for all animals, from single cell yeast to humans, with their drastically different biology, evolutionary history, and complexity; it is rapidly (within few percent of life span) reversible. Such law is unique for live systems with their homeostatic self-adjustment to environment (cf. thermodynamics of liquids and glasses). The law which is valid for all live, and only live, systems is their specific natural law. Mortality is an instrument of natural selection and biological diversity. Its law, which is preserved in evolution of all species, is a conservation law of mortality, selection, evolution, biology. The law implies new kind of intrinsic mortality and adaptation which dominate in evolutionary unprecedented protected populations and, in contrast to species specific natural selection, proceed via universal stepwise rungs and reduce to universal cellular mechanism. The law demonstrates that intrinsic mortality and at least certain aspects of aging are disposable evolutionary byproducts, and directed genetic and/or biological changes may yield healthy and vital Methuselah lifespan. This is consistent with experiments. Universality implies that single cell yeast may provide a master key to the cellular mechanism of universal mortality, aging, selection, evolution, and its regulation in all animals. One may look for its manifestations in animal cells also, e.g., in their replicative senescence and cancer. Evolutionary origin and genetic nature of universality are suggested.

  1. [Liver transplants from living donors].

    PubMed

    Rogiers, X; Danninger, F; Malagó, M; Knoefel, W T; Gundlach, M; Bassas, A; Burdelski, M; Broelsch, C E

    1996-03-01

    In this article the authors discuss the advantages of Living Related Liver Transplantation (LRLT), criteria for the selection of donors and the standard operation technique. Among a total of 241 liver transplantation (LTx), 42 LRLT were performed at the University of Hamburg between October 1, 1991 and December 19, 1994. The body weight of recipients for LRLT ranged from 4,6 to 39 kg, with 64,2% having less than 10 kg. The volume of the donor left lateral liver lobe ranged from 100 cc to 350 cc. The average one year survival rate among electively operated patients-status 3-4 (UNOS 1995 classification) was 86.7%, two year survival rate 83.3%. The main advantages of LRLT are consired the following: 1. Absence of mortality on the waiting list, 2. Optimal timing of the transplantation (elective procedure, patient in a good condition), 3. Excellent organ (no primary non function), 4. A possible immunologic advantage, 5. Relief of the waiting list for cadaveric organs, 6. Psychological benefit for the family, 7. Cost effectiveness. Potential candidates for living donation with more than one cardiovascular risk factors were excluded. Social and psychological reasons leading to rejection of candidates were as follows: unstable family structure, expected professional or financial difficulties after living donation or withdrawal from consent. LRLT gives parents of a child with TLD a chance to avoid the risk of death on the waiting list or primary non function of the graft. LRLT has therefore established an important place in pediatric liver transplantation. PMID:8768973

  2. Exact law of live nature

    NASA Astrophysics Data System (ADS)

    Azbel‧, Mark Ya.

    2005-08-01

    The exact law of mortality dynamics in changing populations and environment is derived. It includes no explicit characteristics of animal-environment interactions (metabolism, etc.) which are a must for life; it is universal for all animals, from single-cell yeast to humans, with their drastically different biology, evolutionary history, and complexity; it is rapidly (within few percent of life span) reversible. Such a law is unique for live systems with their homeostatic self-adjustment to environment (cf. thermodynamics of liquids and glasses). The law which is valid for all live, and only live, systems is their specific natural law. Mortality is an instrument of natural selection and biological diversity. Its law, which is preserved in evolution of all species, is a conservation law of mortality, selection, evolution, biology. The law implies new kinds of intrinsic mortality and adaptation which dominate in evolutionary unprecedented protected populations and, in contrast to species-specific natural selection, proceed via universal stepwise rungs and reduce to universal cellular mechanism. The law demonstrates that intrinsic mortality and at least certain aspects of aging are disposable evolutionary byproducts, and directed genetic and/or biological changes may yield healthy and vital Methuselah lifespan. This is consistent with experiments. Universality implies that single-cell yeast may provide a master key to the cellular mechanism of universal mortality, aging, selection, evolution, and its regulation in all animals. One may look for its manifestations in animal cells also, e.g., in their replicative senescence and cancer. Evolutionary origin and genetic nature of universality are suggested.

  3. Short-Lived Climate Pollution

    NASA Astrophysics Data System (ADS)

    Pierrehumbert, R. T.

    2014-05-01

    Although carbon dioxide emissions are by far the most important mediator of anthropogenic climate disruption, a number of shorter-lived substances with atmospheric lifetimes of under a few decades also contribute significantly to the radiative forcing that drives climate change. In recent years, the argument that early and aggressive mitigation of the emission of these substances or their precursors forms an essential part of any climate protection strategy has gained a considerable following. There is often an implication that such control can in some way make up for the current inaction on carbon dioxide emissions. The prime targets for mitigation, known collectively as short-lived climate pollution (SLCP), are methane, hydrofluo-rocarbons, black carbon, and ozone. A re-examination of the issues shows that the benefits of early SLCP mitigation have been greatly exaggerated, largely because of inadequacies in the methodologies used to compare the climate effects of short-lived substances with those of CO2, which causes nearly irreversible climate change persisting millennia after emissions cease. Eventual mitigation of SLCP can make a useful contribution to climate protection, but there is little to be gained by implementing SLCP mitigation before stringent carbon dioxide controls are in place and have caused annual emissions to approach zero. Any earlier implementation of SLCP mitigation that substitutes to any significant extent for carbon dioxide mitigation will lead to a climate irreversibly warmer than will a strategy with delayed SLCP mitigation. SLCP mitigation does not buy time for implementation of stringent controls on CO2 emissions.

  4. Live from the Moon - Impact!

    NASA Technical Reports Server (NTRS)

    2002-01-01

    On March 24, 1965, a nationwide TV audience watched live video from Ranger 9 as it purposefully crashed into the Moon within the crater Alphonsus. Ranger's six cameras sent back more than 5800 video images during the last 18 minutes of its 3-day journey, the last of the Ranger Project. The last few images show the lunar surface in detail from a few hundred meters above.

    This sequence of images from Camera A was converted from video to film to laser disc to digital files.

  5. Blood Banking in Living Droplets

    PubMed Central

    Shao, Lei; Zhang, Xiaohui; Xu, Feng; Song, YoungSeok; Keles, Hasan Onur; Matloff, Laura; Markel, Jordan; Demirci, Utkan

    2011-01-01

    Blood banking has a broad public health impact influencing millions of lives daily. It could potentially benefit from emerging biopreservation technologies. However, although vitrification has shown advantages over traditional cryopreservation techniques, it has not been incorporated into transfusion medicine mainly due to throughput challenges. Here, we present a scalable method that can vitrify red blood cells in microdroplets. This approach enables the vitrification of large volumes of blood in a short amount of time, and makes it a viable and scalable biotechnology tool for blood cryopreservation. PMID:21412411

  6. Living and Working in Space

    NASA Technical Reports Server (NTRS)

    Roman, Monserrate C.

    2000-01-01

    This document is a presentation about some of the challenges of living and working in space. The presentation shows slides of the Apollo 11 liftoff, Skylab in orbit, a Space Shuttle launch, and a slide of the International Space Station. It reviews the needs and effluents of the astronauts per day, and the Environmental Control and Life Support (ECLS) systems. It shows a flow diagram of the Space Station Regenerative ECLS, which shows the various systems, and how they interact to control the environment and recycle the air, and water. There are other slides some of which show astronauts eating, brushing teeth, shaving, and sipping from a sip bottle while exercising.

  7. Live a Full Life with Fibro

    MedlinePlus

    ... Live a Full Life with Fibro Page Content Fibromyalgia is a chronic pain condition that affects 10 ... family, you can live an active life with fibromyalgia. Talking with Your Physician Take the first step ...

  8. Teasing Out Where the Tokers Live

    MedlinePlus

    ... 160089.html Teasing Out Where the Tokers Live Marijuana use more common in western U.S., report finds To ... States you live in, a new survey suggests. Marijuana use by Americans is highest in the West and ...

  9. Optical trapping inside living organisms

    NASA Astrophysics Data System (ADS)

    Hansen, Poul M.; Oddershede, Lene B.

    2005-08-01

    We use optical tweezers to investigate processes happening inside ving cells. In a previous study, we trapped naturally occurring lipid granules inside living yeast cells, and used them to probe the viscoelastic properties of the cytoplasm. However, we prefer to use probes which can be specifically attached to various organelles within the living cells in order to optically quantify the forces acting on these organelles. Therefore, we have chosen to use nanometer sized gold beads as probes. These gold beads can be conjugated and attached chemically to the organelles of interest. Only Rayleigh metallic particles can be optically trapped and for these it is the case that the larger the beads, the larger the forces which can be exerted and thus measured using optical tweezers. The gold nanoparticles are injected into the cytoplasm using micropipettes. The very rigid cell wall of the S. pombe yeast cells poses a serious obstacle to this injection. In order to be able to punch a hole in the cell, first, the cells have to be turned into protoplasts, where only a lipid bilayer separates the cytoplasm from the surrounding media. We show how to perform micropipette delivery into the protoplasts and also how the protoplasts can be ablated using the trapping laserlight. Finally, we demonstrate that we can transform the protoplasts back to normal yeast cells.

  10. Audio watermarking for live performance

    NASA Astrophysics Data System (ADS)

    Tachibana, Ryuki

    2003-06-01

    Audio watermarking has been used mainly for digitally stored content. Using real-time watermark embedding, its coverage can be extended to live broadcasts and live performances. In general, a conventional embedding algorithm receives a host signal (HS) and outputs the summation of the HS and a watermark signal (WS). However, when applied to real-time embedding, there are two problems: (1) delay of the HS, and (2) possible interruption of the broadcast. To solve these problems, we propose a watermark generation algorithm that outputs only a WS, and a system composition method in which a mixer outside the computer mixes the WS generated by the algorithm and the HS. In addition, we propose a new composition method "sonic watermarking." In this composition method, the sound of the HS and the sound of the WS are played separately by two speakers, and the sounds are mixed in the air. Using this composition method, it would be possible to generate a watermarking sound in a concerto hall so that the watermark could be detected from content recorded by audience members who have recording devices at their seats. We report on the results of experiments and discuss the merits and flaws of various real-time watermarking composition methods.

  11. Live Fire Range Environmental Assessment

    SciTech Connect

    1993-08-01

    The Central Training Academy (CTA) is a DOE Headquarters Organization located in Albuquerque, New Mexico, with the mission to effectively and efficiently educate and train personnel involved in the protection of vital national security interests of DOE. The CTA Live Fire Range (LFR), where most of the firearms and tactical training occurs, is a complex separate from the main campus. The purpose of the proposed action is to expand the LFR to allow more options of implementing required training. The Department of Energy has prepared this Environmental Assessment (EA) for the proposed construction and operation of an expanded Live Fire Range Facility at the Central Training Academy in Albuquerque, New Mexico. Based on the analysis in the EA, DOE has determined that the proposed action is not a major Federal action significantly affecting the quality of the human environment, within the meaning of the National Environmental Policy Act (NEPA) of 1969. Therefore, the preparation of an environmental impact statement is not required and DOE is issuing this Finding of No Significant Impact (FONSI).

  12. Living well in the Neuropolis

    PubMed Central

    Fitzgerald, Des; Rose, Nikolas; Singh, Ilina

    2016-01-01

    Abstract This paper is about the relationship between cities and brains: it charts the back‐and‐forth between the hectic, stressful lives of urban citizens, and a psychological and neurobiological literature that claims to make such stress both visible and knowable. But beyond such genealogical labour, the paper also asks: what can a sociology concerned with the effects of ‘biosocial’ agencies take from a scientific literature on the urban brain? What might sociology even contribute to that literature, in its turn? To investigate these possibilities, the paper centres on the emergence and description of what it calls ‘the Neuropolis’ – a term it deploys to hold together both an intellectual and scientific figure and a real, physical enclosure. The Neuropolis is an image of the city embedded in neuropsychological concepts and histories, but it also describes an embodied set of (sometimes pathological) relations and effects that take places between cities and the people who live in them. At the heart of the paper is an argument that finding a way to thread these phenomena together might open up new paths for thinking about ‘good’ life in the contemporary city. Pushing at this claim, the paper argues that mapping the relations, histories, spaces, and people held together by this term is a vital task for the future of urban sociology. PMID:27397945

  13. Living Free: A Teacher Information Booklet.

    ERIC Educational Resources Information Center

    Mello, Robin

    This workbook helps adolescents learn how to take charge of their own lives and happiness. The underlying idea is to teach them how to live responsibly. By learning to live responsibly, adolescents have the best chance of avoiding drugs, alcohol, and other addictive behaviors such as overeating and overspending. The workbook explains the steps to…

  14. Magnetic Levitational Assembly for Living Material Fabrication.

    PubMed

    Tasoglu, Savas; Yu, Chu Hsiang; Liaudanskaya, Volha; Guven, Sinan; Migliaresi, Claudio; Demirci, Utkan

    2015-07-15

    Functional living materials with microscale compositional topographies are prevalent in nature. However, the creation of biomaterials composed of living micro building blocks, each programmed by composition, functionality, and shape, is still a challenge. A powerful yet simple approach to create living materials using a levitation-based magnetic method is presented. PMID:25872008

  15. Living Arrangement Preferences among Elderly People.

    ERIC Educational Resources Information Center

    Beland, Francois

    1987-01-01

    Compared older adults living alone or with only a spouse to those living with children, relatives, or friends. Found that those living alone or with a spouse were more likely to prefer to move into alternate setting (senior housing or nursing home). Findings revealed limited evidence to support view that residency with others is substitute for…

  16. Magnetic Levitational Assembly for Living Material Fabrication.

    PubMed

    Tasoglu, Savas; Yu, Chu Hsiang; Liaudanskaya, Volha; Guven, Sinan; Migliaresi, Claudio; Demirci, Utkan

    2015-07-15

    Functional living materials with microscale compositional topographies are prevalent in nature. However, the creation of biomaterials composed of living micro building blocks, each programmed by composition, functionality, and shape, is still a challenge. A powerful yet simple approach to create living materials using a levitation-based magnetic method is presented.

  17. Use of Live Supervision in Counselor Preparation.

    ERIC Educational Resources Information Center

    Bubenzer, Donald L.; And Others

    1991-01-01

    Investigated live supervision in counselor preparation programs by surveying 307 counselor preparation programs. Live supervision was used at 157 institutions and was used in preparing individual, group, and marriage and family counselors. At least 75 percent of programs provided live supervision weekly. Techniques of cotherapy and remote viewing…

  18. The Cassini Live Update Process

    NASA Technical Reports Server (NTRS)

    Vandermey, Nancy; Ray, Trina; Wallis, Brad; Roumeliotis, Chris

    2008-01-01

    The Cassini orbiter is an international science mission to the Saturnian system with 12 science instruments onboard. The Cassini spacecraft lacks a scan platform, which means the entire spacecraft must be rotated to control pointing of any one instrument's boresight. The resulting complex sequences of commands are built beginning many months before execution onboard. Late ephemeris updates from improved navigation data (i.e. after an orbital trim maneuver) often result in pointing commands in the sequence no longer being accurate enough to obtain the desired science observation. This paper will provide an overview of how Cassini uses live updates to address this potential loss of data, including the software developed for this process.

  19. Molecular Spectroscopy of Living Systems

    NASA Astrophysics Data System (ADS)

    Cheng, Ji-Xin

    2016-06-01

    Molecular spectroscopy has been a powerful tool in the study of molecules in gas phase, condensed phase, and at interfaces. The transition from in vitro spectroscopy to spectroscopic imaging of living systems is opening new opportunities to reveal cellular machinery and to enable molecule-based diagnosis (Science 2015, 350: 1054). Such a transition involves more than a simple combination of spectrometry and microscopy. In this presentation, I will discuss the most recent efforts that have pushed the physical limits of spectroscopic imaging in terms of spectral acquisition speed, detection sensitivity, spatial resolution and imaging depth. I will further highlight significant applications in functional analysis of single cells and in label-free detection of diseases.

  20. ATLAS Live: Collaborative Information Streams

    NASA Astrophysics Data System (ADS)

    Goldfarb, Steven; ATLAS Collaboration

    2011-12-01

    I report on a pilot project launched in 2010 focusing on facilitating communication and information exchange within the ATLAS Collaboration, through the combination of digital signage software and webcasting. The project, called ATLAS Live, implements video streams of information, ranging from detailed detector and data status to educational and outreach material. The content, including text, images, video and audio, is collected, visualised and scheduled using digital signage software. The system is robust and flexible, utilizing scripts to input data from remote sources, such as the CERN Document Server, Indico, or any available URL, and to integrate these sources into professional-quality streams, including text scrolling, transition effects, inter and intra-screen divisibility. Information is published via the encoding and webcasting of standard video streams, viewable on all common platforms, using a web browser or other common video tool. Authorisation is enforced at the level of the streaming and at the web portals, using the CERN SSO system.

  1. Towards a living earth simulator

    NASA Astrophysics Data System (ADS)

    Paolucci, M.; Kossman, D.; Conte, R.; Lukowicz, P.; Argyrakis, P.; Blandford, A.; Bonelli, G.; Anderson, S.; de Freitas, S.; Edmonds, B.; Gilbert, N.; Gross, M.; Kohlhammer, J.; Koumoutsakos, P.; Krause, A.; Linnér, B.-O.; Slusallek, P.; Sorkine, O.; Sumner, R. W.; Helbing, D.

    2012-11-01

    The Living Earth Simulator (LES) is one of the core components of the FuturICT architecture. It will work as a federation of methods, tools, techniques and facilities supporting all of the FuturICT simulation-related activities to allow and encourage interactive exploration and understanding of societal issues. Society-relevant problems will be targeted by leaning on approaches based on complex systems theories and data science in tight interaction with the other components of FuturICT. The LES will evaluate and provide answers to real-world questions by taking into account multiple scenarios. It will build on present approaches such as agent-based simulation and modeling, multiscale modelling, statistical inference, and data mining, moving beyond disciplinary borders to achieve a new perspective on complex social systems.

  2. Adolescents' lived experience of epilepsy.

    PubMed

    Eklund, Pernilla Garmy; Sivberg, Bengt

    2003-02-01

    To improve the well-being of adolescents with epilepsy, research is needed on how adolescents cope. In this study, Lazarus' model of stress and coping and Antonovsky's Theory of Sense of Coherence were used as the theoretical framework. The aim was to describe the lived experience of adolescents with epilepsy and their coping skills. The participants were 13-19 years old with an epilepsy diagnosis but without mental retardation or cerebral palsy. The study was performed in southern Sweden at the pediatric department of a university hospital. Semistructured and open-ended interviews were conducted with 13 adolescents. The transcripts were analyzed with manifest and latent content analysis. All the adolescents had developed strategies to cope with the emotional strains caused by epilepsy. They experienced strains from the seizures, limitation of leisure activities, side effects of medication, and feelings of being different. The coping strategies described were finding support, being in control, and experimenting.

  3. The experience of living with persistent pain.

    PubMed

    Carson, M G; Mitchell, G J

    1998-12-01

    This article reports themes from a descriptive exploratory study conducted with 17 persons who live with persistent pain. The purpose of the research was to enhance understanding of what it is like to live with persistent pain from the perspectives of those who live the experience. Participant narratives of living with persistent pain included descriptions of physical pain as well as the pain of loss and restriction. The theory of human becoming provided the framework for guiding researcher-participant discussions and interpretation of findings. Three themes arose from the analysis: forbearance surfaces with the drain of persistent anguish; isolating retreats coexist with comforting engagements; and hope for relief clarifies priorities for daily living. The findings expand understanding about quality of life for persons who live with persistent pain. They also challenge nurses to begin focusing on the person who lives with pain as the messenger of how to endure. Recommendations, based on new understanding, are offered for practice and additional research.

  4. Living science: Science as an activity of living beings.

    PubMed

    MacLennan, Bruce J

    2015-12-01

    The philosophy of science should accommodate itself to the facts of human existence, using all aspects of human experience to adapt more effectively, as individuals, species, and global ecosystem. This has several implications: (1) Our nature as sentient beings interacting with other sentient beings requires the use of phenomenological methods to investigate consciousness. (2) Our embodied, situated, purposeful physical interactions with the world are the foundation of scientific understanding. (3) Aristotle's four causes are essential for understanding living systems and, in particular, the final cause aids understanding the role of humankind, and especially science, in the global ecosystem. (4) In order to fulfill this role well, scientists need to employ the full panoply of human faculties. These include the consciousness faculties (thinking, sensation, feeling, intuition), and therefore, as advocated by many famous scientists, we should cultivate our aesthetic sense, emotions, imagination, and intuition. Our unconscious faculties include archetypal structures common to all humans, which can guide scientific discovery. By striving to engage the whole of human nature, science will fulfill better its function for humans and the global ecosystem.

  5. Dynamical Signatures of Living Systems

    NASA Technical Reports Server (NTRS)

    Zak, M.

    1999-01-01

    One of the main challenges in modeling living systems is to distinguish a random walk of physical origin (for instance, Brownian motions) from those of biological origin and that will constitute the starting point of the proposed approach. As conjectured, the biological random walk must be nonlinear. Indeed, any stochastic Markov process can be described by linear Fokker-Planck equation (or its discretized version), only that type of process has been observed in the inanimate world. However, all such processes always converge to a stable (ergodic or periodic) state, i.e., to the states of a lower complexity and high entropy. At the same time, the evolution of living systems directed toward a higher level of complexity if complexity is associated with a number of structural variations. The simplest way to mimic such a tendency is to incorporate a nonlinearity into the random walk; then the probability evolution will attain the features of diffusion equation: the formation and dissipation of shock waves initiated by small shallow wave disturbances. As a result, the evolution never "dies:" it produces new different configurations which are accompanied by an increase or decrease of entropy (the decrease takes place during formation of shock waves, the increase-during their dissipation). In other words, the evolution can be directed "against the second law of thermodynamics" by forming patterns outside of equilibrium in the probability space. Due to that, a specie is not locked up in a certain pattern of behavior: it still can perform a variety of motions, and only the statistics of these motions is constrained by this pattern. It should be emphasized that such a "twist" is based upon the concept of reflection, i.e., the existence of the self-image (adopted from psychology). The model consists of a generator of stochastic processes which represents the motor dynamics in the form of nonlinear random walks, and a simulator of the nonlinear version of the diffusion

  6. Access to care save lives.

    PubMed

    Blaney, C L

    1994-02-01

    Emergency treatment of such major complications of pregnancy as obstructed labor, hemorrhage, infection, hypertension disorders, and the effects of unsafe abortion, helps ameliorate morbidity and prevent mortality. Access to life-saving treatment (e.g., antibiotics, Cesarean sections, and blood transfusions) in developing countries is limited. Maternal mortality in one area of The Gambia, for example, is 2200 per 100,000 births. Improving access to care depends upon the availability of these services in communities, trained health personnel, service improvements, transportation provision, and community education. Detection of complications and early referral to an appropriate facility with a supportive and professional environment is key to saving lives. Political will and public pressure are needed before improvement in services can be successfully accomplished; politicians may ignore women with low status. Barriers to care are physical, cultural, technical, and economic. Cost or distance from home may prevent women from seeking care. Infection, hemorrhage, and uterine injury are frequently related to unsafe abortions, particularly among teenage women. Hospitals must be equipped with a reliable management system, surgical facilities, and clinical services. The WHO recommends upgrading community health centers with trained personnel, adequate supervision, and equipment. In Uganda, midwives are specially trained in advanced skills for use in remote areas: administration of oxytocin to evacuate the uterus and reduce bleeding, use of antibiotics for infections, and surgical repair of vaginal tears. Nurses in Zaire are trained to do Cesarean sections. In Sierra Leone and Nigeria, doctors are encouraged to receive training in obstetrics and to be posted in rural areas. In Sierra Leone, young men are trained to bring pregnant women in to care on stretchers. Maternity waiting homes near hospitals are another means to save lives. Lack of permission from a male relative may

  7. Face liveness detection using defocus.

    PubMed

    Kim, Sooyeon; Ban, Yuseok; Lee, Sangyoun

    2015-01-01

    In order to develop security systems for identity authentication, face recognition (FR) technology has been applied. One of the main problems of applying FR technology is that the systems are especially vulnerable to attacks with spoofing faces (e.g., 2D pictures). To defend from these attacks and to enhance the reliability of FR systems, many anti-spoofing approaches have been recently developed. In this paper, we propose a method for face liveness detection using the effect of defocus. From two images sequentially taken at different focuses, three features, focus, power histogram and gradient location and orientation histogram (GLOH), are extracted. Afterwards, we detect forged faces through the feature-level fusion approach. For reliable performance verification, we develop two databases with a handheld digital camera and a webcam. The proposed method achieves a 3.29% half total error rate (HTER) at a given depth of field (DoF) and can be extended to camera-equipped devices, like smartphones. PMID:25594594

  8. Live your life out loud.

    PubMed

    Gatrell, Nanette; Rothblum, Esther

    2014-01-01

    Susan Love, MD, MBA, has dedicated her professional life to the eradication of breast cancer. Author of the bestselling Dr. Susan Love's Breast Book, Dr. Susan Love's Menopause and Hormone Book, and Live a Little, Susan is Chief Visionary Officer of the Dr. Susan Love Research Foundation where she oversees an active research program centered on breast cancer cause and prevention. Susan co-founded the National Breast Cancer Coalition in the early 1990s, and she served on President Clinton's National Cancer Advisory Board from 1998-2004. Her recent projects include recruiting 377,000 women for the Love Army of Women, an Internet program that partners women and scientists to accelerate breast cancer research; and the online Health of Women Study designed to identify the cause of breast cancer. A recipient of six honorary doctorate degrees, Susan is Clinical Professor of Surgery at the David Geffen School of Medicine at the University of California, Los Angeles. In June of 2012, Susan was diagnosed with acute myelogenous leukemia and was treated with an allogenic stem cell transplant.

  9. Face Liveness Detection Using Defocus

    PubMed Central

    Kim, Sooyeon; Ban, Yuseok; Lee, Sangyoun

    2015-01-01

    In order to develop security systems for identity authentication, face recognition (FR) technology has been applied. One of the main problems of applying FR technology is that the systems are especially vulnerable to attacks with spoofing faces (e.g., 2D pictures). To defend from these attacks and to enhance the reliability of FR systems, many anti-spoofing approaches have been recently developed. In this paper, we propose a method for face liveness detection using the effect of defocus. From two images sequentially taken at different focuses, three features, focus, power histogram and gradient location and orientation histogram (GLOH), are extracted. Afterwards, we detect forged faces through the feature-level fusion approach. For reliable performance verification, we develop two databases with a handheld digital camera and a webcam. The proposed method achieves a 3.29% half total error rate (HTER) at a given depth of field (DoF) and can be extended to camera-equipped devices, like smartphones. PMID:25594594

  10. Live attenuated intranasal influenza vaccine.

    PubMed

    Esposito, Susanna; Montinaro, Valentina; Groppali, Elena; Tenconi, Rossana; Semino, Margherita; Principi, Nicola

    2012-01-01

    Annual vaccination is the most effective means of preventing and controlling influenza epidemics, and the traditional trivalent inactivated vaccine (TIV) is by far the most widely used. Unfortunately, it has a number of limitations, the most important of which is its poor immunogenicity in younger children and the elderly, the populations at greatest risk of severe influenza. Live attenuated influenza vaccine (LAIV) has characteristics that can overcome some of these limitations. It does not have to be injected because it is administered intranasally. It is very effective in children and adolescents, among whom it prevents significantly more cases of influenza than the traditional TIV. However, its efficacy in adults has not been adequately documented, which is why it has not been licensed for use by adults by the European health authorities. LAIV is safe and well tolerated by children aged > 2 y and adults, but some concerns arisen regarding its safety in younger children and subjects with previous asthma or with recurrent wheezing. Further studies are needed to solve these problems and to evaluate the possible role of LAIV in the annual vaccination of the general population.

  11. Living "anyway:" stories of access.

    PubMed

    Dykewomon, Elana

    2014-01-01

    Elana Dykewomon's 1974 novel, Riverfinger Women, was among the first lesbian books with a "happy ending." Her seven books of fiction and poetry include the Lambda Award winner Beyond the Pale (now an audio and e-book) and Lambda nominee, Risk. She was an editor of the lesbian-feminist journal, Sinister Wisdom, for eight years. Her literary work foregrounds the lesbian heroic as integral to women's communities. As a social justice activist, she has organized and participated in anti-war, anti-racist, anti-classist, fat and disability rights work since the 1970s. She is now working with Old Lesbians Organizing for Change. She is happy to live embedded in dyke community as a lesbian radical committed to a loving justice. While she suffered psychiatric abuse at 13 (and acknowledges long-term adaptive behavior on that account), she has not experienced disabling mental illness since. Her primary disabilities are mobility impairment through severe, progressive arthritis and constant low-to-powerful pain, sometimes diagnosed as fibromyalgia. Her acute illnesses include pancreatitis and a rare-in-adults kidney disease currently in remission. PMID:24400626

  12. Three dimensional living neural networks

    NASA Astrophysics Data System (ADS)

    Linnenberger, Anna; McLeod, Robert R.; Basta, Tamara; Stowell, Michael H. B.

    2015-08-01

    We investigate holographic optical tweezing combined with step-and-repeat maskless projection micro-stereolithography for fine control of 3D positioning of living cells within a 3D microstructured hydrogel grid. Samples were fabricated using three different cell lines; PC12, NT2/D1 and iPSC. PC12 cells are a rat cell line capable of differentiation into neuron-like cells NT2/D1 cells are a human cell line that exhibit biochemical and developmental properties similar to that of an early embryo and when exposed to retinoic acid the cells differentiate into human neurons useful for studies of human neurological disease. Finally induced pluripotent stem cells (iPSC) were utilized with the goal of future studies of neural networks fabricated from human iPSC derived neurons. Cells are positioned in the monomer solution with holographic optical tweezers at 1064 nm and then are encapsulated by photopolymerization of polyethylene glycol (PEG) hydrogels formed by thiol-ene photo-click chemistry via projection of a 512x512 spatial light modulator (SLM) illuminated at 405 nm. Fabricated samples are incubated in differentiation media such that cells cease to divide and begin to form axons or axon-like structures. By controlling the position of the cells within the encapsulating hydrogel structure the formation of the neural circuits is controlled. The samples fabricated with this system are a useful model for future studies of neural circuit formation, neurological disease, cellular communication, plasticity, and repair mechanisms.

  13. Perspective on live varicella vaccine.

    PubMed

    Gershon, Anne A; Katz, Samuel L

    2008-03-01

    The attenuation of varicella-zoster virus (VZV) by Takahashi in 1974 was a remarkable achievement. It swiftly led to development of a live vaccine against chickenpox, which was initially tested in Japan. With its successful employment in immunocompromised children to prevent morbidity and mortality due to varicella, the vaccine began to be tested in healthy children in Japan and elsewhere. In the United States, vaccine use progressed from extensive clinical trials that demonstrated safety and efficacy to universal immunization of healthy infants and children. In the past 10 years, >30 million healthy American individuals, mostly children, have been vaccinated. With increasing use of vaccine, there has been a concomitant decrease in the incidence of disease, along with decreases in hospitalizations and deaths due to VZV. To improve protection, however, a 2-dose schedule of immunization was recommended for routine use in all children by the Centers for Disease Control and Prevention in June 2006. At roughly the same time, licensure of the combined measles-mumps-rubella-varicella vaccine was completed, which allowed harmonization of immunization against these 4 viruses with 1 injection given twice in childhood. Concomitantly, a version of the varicella vaccine with 10 times the titer was developed for immunization of healthy individuals >60 years of age against herpes zoster (HZ). Although elimination of VZV from human populations may not yet be possible, the combined approach of immunization against both varicella in childhood and HZ in adulthood in the developed world are predicted to dramatically increase our control of this troublesome virus.

  14. ``Backpack'' Functionalized Living Immune Cells

    NASA Astrophysics Data System (ADS)

    Swiston, Albert; Um, Soong Ho; Irvine, Darrell; Cohen, Robert; Rubner, Michael

    2009-03-01

    We demonstrate that functional polymeric ``backpacks'' built from polyelectrolyte multilayers (PEMs) can be attached to a fraction of the surface area of living, individual lymphocytes. Backpacks containing fluorescent polymers, superparamagnetic nanoparticles, and commercially available quantum dots have been attached to B and T-cells, which may be spatially manipulated using a magnetic field. Since the backpack does not occlude the entire cellular surface from the environment, this technique allows functional synthetic payloads to be attached to a cell that is free to perform its native functions, thereby synergistically utilizing both biological and synthetic functionalities. For instance, we have shown that backpack-modified T-cells are able to migrate on surfaces for several hours following backpack attachment. Possible payloads within the PEM backpack include drugs, vaccine antigens, thermally responsive polymers, nanoparticles, and imaging agents. We will discuss how this approach has broad potential for applications in bioimaging, single-cell functionalization, immune system and tissue engineering, and cell-based therapeutics where cell-environment interactions are critical.

  15. "Living theatre, theatre of life".

    PubMed

    Wenzel, E

    1987-09-01

    Young people love to play theatre--in one way or another. They like to play with behaviours, costumes, words, communication patterns, etc.; they like to disguise themselves, to create certain spheres and scenes of drama and tragedy, excitement and extacy, satire and irony, morals and decadence. Due to the particular uncertainties of the adolescent passage, youth oscillates between taking life both, too seriously and easy. Searching for identity and integration, they tend to experiment with styles of behaviour and culturally defined patterns of lifestyles conductive to well-being. Sometimes, life is perceived as pure entertainment, and sometimes as pure drama. It's living theatre and theatre of life. On the one hand it is "acting out", on the other hand it is playing precisely defined roles. And, in-between, it is always the question: Who am I? They tend to slip into roles in order to check out whether they are willing to accept their implications with regard to the priorities they have set so far.

  16. Nanocapsules: coating for living cells.

    PubMed

    Krol, Silke; Diaspro, Alberto; Magrassi, Raffaella; Ballario, Paola; Grimaldi, Benedetto; Filetici, Patrizia; Ornaghi, Prisca; Ramoino, Paola; Gliozzi, Alessandra

    2004-03-01

    One of the most promising tools for future applications in science and medicine is the use of nanotechnologies. Especially self-assembly systems, e.g., polyelectrolyte (PE) capsules prepared by means of the layer-by-layer technique with tailored properties, fulfill the requirements for nano-organized systems in a satisfactory manner. The nano-organized shells are suitable as coating for living cells or artificial tissue to prevent immune response. With these shells, material can be delivered to predefined organs. In this paper, some preliminary results are presented, giving a broad overview over the possibilities to use nano-organized capsules. Based on the observations that the cells while duplicating break the capsule a mutant yeast strain (Saccharomyces cerevisiae), which express GFP-tubulin under galactose promotion, was investigated by means of confocal laser scanning microscopy. The measurements reveal an increased surface charge in the region of buds developed prior encapsulation. In order to test the used PE pair for cytotoxicity, germinating conidia of the fungi Neurospora crassa were coated. The investigation with fluorescence microscopy shows a variation in the surface charge for the growing region and the conidium poles. The capsules exhibit interesting properties as valuable tool in science and a promising candidate for application in the field of medicine.

  17. Living musical instruments and inanimate body parts?

    PubMed

    Barbarotto, R; Capitani, E; Laiacona, M

    2001-01-01

    In the literature about category effects in semantic memory, body parts and musical instruments are often considered atypical, because in cases with a disproportionate impairment of living categories body parts are relatively spared, while musical instruments are often severely defective. In this study the performance of 57 subjects affected by diseases generally associated with lexical-semantic impairment, for the most part Alzheimer's disease and other forms of cortical degeneration, but also herpetic encephalitis and traumatic brain damage are analyzed. The subjects were given a picture naming task tapping eight categories: three living categories (animals, fruits and vegetables) and three non-living categories (tools, furniture and vehicles), plus body parts and musical instruments. On a preliminary analysis at the group level, body parts were the least impaired category and musical instruments the most severely impaired, the six living and non-living categories being intermediate. However, these differences disappeared after covariance for lexical frequency, name agreement and age of acquisition. The relationship between living categories, non-living categories, musical instruments and body parts was investigated by means of a Lisrel model of Confirmatory Factor Analysis. Two latent variables related to living and non-living categories respectively were defined, and it was found that both body parts and musical instruments were significantly related only with non-living categories. The results showed that the definition of the latent variable expressing the substrate of non-living categories was less satisfactory than that expressing the living categories. On this basis, the conclusions of this study appear statistically definite but their psychological interpretation is less straightforward.

  18. Live Blogging Science News: The Rosetta Mission

    NASA Astrophysics Data System (ADS)

    Clark, S.

    2016-03-01

    When one of the world's most popular online news websites decides to cover a space science event live, you know that something big is brewing. Stuart Clark reports on how live blogging can be used for science reporting and how an idea that was triggered by his observations during the Rosetta flyby of the asteroid Lutetia and the landing of the Curiosity rover on Mars led to him live blogging two of Rosetta's most memorable occasions for The Guardian newspaper.

  19. Only One Earth: Living for the Future.

    ERIC Educational Resources Information Center

    Timberlake, Lloyd

    Depicting how people in nine countries can improve their lives while living within their environmental means is the primary aim of this book. Chapter 1 highlights the division of earth's people into the haves and the have nots and describes the inequity of so few having so much of the available resources. Chapter 2 examines the problem of how…

  20. Living with Learning Difficulties: Emma's Story

    ERIC Educational Resources Information Center

    Manners, Paula Jean; Carruthers, Emma

    2006-01-01

    This article is about Emma's experience of living with learning difficulties. Emma expresses a lot of anger, and talks about feelings of loss. This article is interesting to people with learning disabilities because they can see if their experience is like Emma's in any way. This paper presents Emma's story: her experience of living with learning…

  1. [Cultivation of pathogenic free-living amoebae].

    PubMed

    Peng, Heng; Zhu, Huai-Min

    2009-08-01

    The isolation and culture of pathogenic free-living amoebae are useful in the diagnosis and research. This review focuses on the methods of isolation and cultivation of pathogenic free-living amoebae, including sample treatment, culture conditions, passage culture, pathogen detection, and maintenance.

  2. Opening Minds: Using Language to Change Lives

    ERIC Educational Resources Information Center

    Johnston, Peter H.

    2012-01-01

    Sometimes a single word changes everything. In his groundbreaking book "Choice Words", Peter Johnston demonstrated how the things teachers say (and don't say) have surprising consequences for the literate lives of students. Now, in "Opening Minds: Using Language to Change Lives", Peter shows how the words teachers choose affect the worlds students…

  3. 38 CFR 21.76 - Independent living.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...) VOCATIONAL REHABILITATION AND EDUCATION Vocational Rehabilitation and Employment Under 38 U.S.C. Chapter 31 Duration of Rehabilitation Programs § 21.76 Independent living. (a) General. A program of independent living services may be authorized to enable the veteran to: (1) Reach the goals of the program, and...

  4. NASA LIVE Creating a Global Classroom

    ERIC Educational Resources Information Center

    Townes-Young, Katrina L.; Ewing, Virginia R.

    2005-01-01

    This article describes NASA LIVE (Learning through Interactive Videoconferencing Experiences), a free series of videoconferencing programs produced by NASA's Langley Center for Distance Learning in Hampton, Virginia. NASA LIVE is designed for K-12 educators and students, allowing teachers and students to interact with NASA experts in a virtual…

  5. Techniques for Daily Living: Curriculum Guides.

    ERIC Educational Resources Information Center

    Wooldridge, Lillian; And Others

    Presented are specific guides concerning techniques for daily living which were developed by the child care staff at the Illinois Braille and Sight Saving School. The guides are designed for cottage parents of the children, who may have both visual and other handicaps, and show what daily living skills are necessary and appropriate for the…

  6. Inquiry and Living History, Part II

    ERIC Educational Resources Information Center

    Coatney, Sharon; Smalley, Rachel

    2006-01-01

    In the first part of this article, the authors introduced the living history program. This yearly, weeklong program features living portrayals of famous people, which becomes a catalyst for teaching curricular standards, as well as providing the spark for inquiry. Successful implementation of this program requires providing teachers with…

  7. The Experience of Living Kidney Donors

    ERIC Educational Resources Information Center

    Brown, Judith Belle; Karley, Mary Lou; Boudville, Neil; Bullas, Ruth; Garg, Amit X.; Muirhead, Norman

    2008-01-01

    This article describes the experiences, feelings, and ideas of living kidney donors. Using a phenomenological, qualitative research approach, the authors interviewed 12 purposefully selected living kidney donors (eight men and four women), who were between four and 29 years since donation. Interviews were audiotaped, and transcribed verbatim, and…

  8. The Family: A School for Living

    ERIC Educational Resources Information Center

    Garbarino, James

    1976-01-01

    In the face of so many trends toward fragmentation, overspecialization, and instrumentally impersonal relationships in the day-to-day lives of families, the cause of social habitability is best served when we are able to see clearly the ecology of human development: families as schools for living and schools as families. (Author)

  9. Living Skills as a Core Curriculum Component.

    ERIC Educational Resources Information Center

    Dufty, David

    Schools should help students develop daily living skills in addition to basic cognitive skills such as reading, writing, and figuring. Living skills are interpreted to include those skills which help students cope with rapid social change. Skills need to be taught on health and nutrition, safety and first aid, interpersonal relationships, family…

  10. An Interstate Cost-of-Living Index.

    ERIC Educational Resources Information Center

    Nelson, F. Howard

    An interstate cost-of-living index from SMSA-level market basket data is generated in this research. Models are developed for each of the four regions--Northwest, North Central, South, and West--to explain differences in the cost of living among more than 180 SMSAs. Weighted regression coefficients were combined with comparable state-level data to…

  11. Environmental Living Program, Book 1: What's Happening.

    ERIC Educational Resources Information Center

    Baldi, Mary Lou

    This booklet documents, in words and pictures, the Environmental Living Program. This program, which has been in operation since 1969, provides overnight living experiences for elementary and secondary school students at cultural, historic, or prehistoric sites. The sites are National and State parks and private sites in California and Arizona.…

  12. Beauty in the Context of Particular Lives

    ERIC Educational Resources Information Center

    Rautio, Pauliina

    2010-01-01

    This paper is based on empirical research by the author into the everyday lives of people living in a small village. Everyday life is approached as a subjective process in time and space, experienced by particular people in a particular environmental and social context. The data of this research has been collected through correspondence in which…

  13. Bush Blitz TeachLive 2015

    ERIC Educational Resources Information Center

    Greene, Chris

    2015-01-01

    In July 2015, the Bush Blitz TeachLive expedition took place on the Olkola people's traditional lands on Cape York Peninsula in Queensland. Five teachers joined more than a dozen scientists and the Olkola Indigenous rangers to act as research assistants, as part of the eight-day Bush Blitz TeachLive experience. As a team, they discovered new…

  14. Community Living Skills Guide: Looking Good: Hygiene.

    ERIC Educational Resources Information Center

    Kreps, Alice Roelofs; Dreith, Rita Vallero

    One of twenty course guides in the Community Living Skills Guide for the College for Living series, this document provides guidelines and workbook activities for the course, Looking Good: Hygiene. The series of courses for developmentally disabled adults is intended to supplement residential programs and to aid in orienting institutionalized…

  15. Living Citizenship: Transcending the Cultural Divide

    ERIC Educational Resources Information Center

    Coombs, Steven; Potts, Mark

    2013-01-01

    Living citizenship emerging from reflection on an international educational partnership makes a unique contribution to the field and importantly fulfils the British Educational Research Association aim of improving educational practice for the public benefit. This paper explores the conceptual framework of 'living citizenship' as a means…

  16. GIS Live and Web Problem Solving

    ERIC Educational Resources Information Center

    Hagevik, R.; Hales, D.; Harrell, J.

    2007-01-01

    GIS Live is a live, interactive, web problem-solving (WPS) program that partners Geographic Information Systems (GIS) professionals with educators to implement geospatial technologies as curriculum-learning tools. It is a collaborative effort of many government agencies, educational institutions, and professional organizations. Problem-based…

  17. Community Living Skills Guide: Beginning Woodworking.

    ERIC Educational Resources Information Center

    Miller, Steve; Smith, Don

    This is one of twenty course guides in the Community Living Skills Guide for the College for Living series which provides guidelines and workbook activities for the course, Beginning Woodworking, Use of Basic Hand Tools and Shop Safety. The series of courses for developmentally disabled adults is intended to supplement residential programs and to…

  18. Women's Guide to Overseas Living. Second Edition.

    ERIC Educational Resources Information Center

    Piet-Pelon, Nancy J.; Hornby, Barbara

    This book examines issues critical to women and their families who go abroad to live. In advising how to cope effectively with the problems that arise, the book illuminates the advantages of living overseas and offers practical suggestions and guidelines that help women take advantage of the opportunity to share in another culture. Divided into 13…

  19. Reflection on Lived Experience in Educational Research

    ERIC Educational Resources Information Center

    Barnacle, Robyn

    2004-01-01

    While debate about the meaning of hermeneutics and phenomenology for educational research continues, the notion of lived experience, and its application to reflective practice, has become a feature of much that goes by the name of phenomenological within this area. The prevalence of the lived experience model can be attributed in large part to the…

  20. The Independent Living Behavior Checklist: Experimental Edition.

    ERIC Educational Resources Information Center

    Walls, Richard T.; And Others

    The document describes independent living skills, and provides information on how they can be measured. It is explained in an introductory chapter that the checklist is an extensive list of 343 independent living skill objectives specified in terms of conditions (antecedents or givens), behaviors, and standards. Objectives are classified and…

  1. Bringing Art to Life through Living Paintings

    ERIC Educational Resources Information Center

    Stillwagon, Joanne

    2005-01-01

    In this article, the author relates how she and other art teachers developed a "living painting" lesson, an art lesson that incorporates art history and painting. The lesson was developed also in part to help new student-teachers plan a memorable lesson. With the living painting lesson, students will have to choose and recreate famous paintings…

  2. Nanobiomechanics of living cells: a review

    PubMed Central

    Chen, Jinju

    2014-01-01

    Nanobiomechanics of living cells is very important to understand cell–materials interactions. This would potentially help to optimize the surface design of the implanted materials and scaffold materials for tissue engineering. The nanoindentation techniques enable quantifying nanobiomechanics of living cells, with flexibility of using indenters of different geometries. However, the data interpretation for nanoindentation of living cells is often difficult. Despite abundant experimental data reported on nanobiomechanics of living cells, there is a lack of comprehensive discussion on testing with different tip geometries, and the associated mechanical models that enable extracting the mechanical properties of living cells. Therefore, this paper discusses the strategy of selecting the right type of indenter tips and the corresponding mechanical models at given test conditions. PMID:24748952

  3. Financial considerations in living organ donation.

    PubMed

    Jacobs, Cheryl; Thomas, Charlie

    2003-06-01

    The shortage of cadaveric organs and increased success of living donor transplantation support the use of living organ donors. Clinical social workers have the opportunity to explore a variety of donor-specific issues when performing psychosocial evaluations of living donors, including motivation, psychological stability, and personal and family consequences of donation, as well as the direct and indirect financial consequences faced by living donors. Although most donor-related medical costs are covered, other associated expenses are not reimbursable and may put donors at risk for financial hardship. Out-of-pocket expenses also serve as a disincentive to donate for some volunteers. During the evaluation process, healthcare professionals should openly discuss how surgery, recovery, and any potential complications might impact prospective donors' financial situation. Donors can then decide whether they are able to realistically handle the costs of donation. We present the financial dilemmas experienced by many living donors and highlight efforts that have been made to deal with them.

  4. Living will status and desire for living will help among rural Alabama veterans.

    PubMed

    Mahaney-Price, Ann F; Hilgeman, Michelle M; Davis, Lori L; McNeal, Sandre F; Conner, Charles M; Allen, Rebecca S

    2014-10-01

    The purpose of this secondary analysis of data from an earlier intervention study to increase Veterans Administration health care enrollment in rural Alabama veterans was to determine the veterans' living will status, desire for help completing a living will, and relationships between these and demographic, health insurance, health self-report, cumulative illness, disability, and trust characteristics. Baseline data for 201 rural Alabama veterans were extracted from the larger study. Chi-square and t tests were used to analyze group differences in categorical and continuous variables. Logistic regression models were used to determine multivariate associations of variables with living will status and desire for help. Only 13% of participants had living wills. Of those without living wills, 40% expressed a desire for help completing a living will. African Americans were less likely to have living wills than were Caucasians. Participants with more than high school education were more likely to desire help completing living wills than were those with less education. With the exception of moderate-severe respiratory illness, moderate-severe illness was not associated with having a living will. With the exception of moderate-severe vascular illness, moderate-severe illness was not associated with desire for help completing a living will. The racial and educational disparities in living will status and desire for help and the number of participants who desired help completing a living will suggests a need for action to increase advance care planning among rural veterans.

  5. Development of a High-Yield Live Attenuated H7N9 Influenza Virus Vaccine That Provides Protection against Homologous and Heterologous H7 Wild-Type Viruses in Ferrets

    PubMed Central

    Baz, Mariana; Lu, Janine; Paskel, Myeisha; Santos, Celia; Subbarao, Kanta; Jin, Hong; Matsuoka, Yumiko

    2014-01-01

    ABSTRACT Live attenuated H7N9 influenza vaccine viruses that possess the hemagglutinin (HA) and neuraminidase (NA) gene segments from the newly emerged wild-type (wt) A/Anhui/1/2013 (H7N9) and six internal protein gene segments from the cold-adapted influenza virus A/Ann Arbor/6/60 (AA ca) were generated by reverse genetics. The reassortant virus containing the original wt A/Anhui/1/2013 HA and NA sequences replicated poorly in eggs. Multiple variants with amino acid substitutions in the HA head domain that improved viral growth were identified by viral passage in eggs and MDCK cells. The selected vaccine virus containing two amino acid changes (N133D/G198E) in the HA improved viral titer by more than 10-fold (reached a titer of 108.6 fluorescent focus units/ml) without affecting viral antigenicity. Introduction of these amino acid changes into an H7N9 PR8 reassortant virus also significantly improved viral titers and HA protein yield in eggs. The H7N9 ca vaccine virus was immunogenic in ferrets. A single dose of vaccine conferred complete protection of ferrets from homologous wt A/Anhui/1/2013 (H7N9) and nearly complete protection from heterologous wt A/Netherlands/219/2003 (H7N7) challenge infection. Therefore, this H7N9 live attenuated influenza vaccine (LAIV) candidate has been selected for vaccine manufacture and clinical evaluation to protect humans from wt H7N9 virus infection. IMPORTANCE In response to the recent avian H7N9 influenza virus infection in humans, we developed a live attenuated H7N9 influenza vaccine (LAIV) with two amino acid substitutions in the viral HA protein that improved vaccine yield by 10-fold in chicken embryonated eggs, the substrate for vaccine manufacture. The two amino acids also improved the antigen yield for inactivated H7N9 vaccines, demonstrating that this finding could great facilitate the efficiency of H7N9 vaccine manufacture. The candidate H7N9 LAIV was immunogenic and protected ferrets against homologous and heterologous

  6. Living Donor Kidney Transplantation: Facilitating Education about Live Kidney Donation--Recommendations from a Consensus Conference.

    PubMed

    Tan, Jane C; Gordon, Elisa J; Dew, Mary Amanda; LaPointe Rudow, Dianne; Steiner, Robert W; Woodle, E Steve; Hays, Rebecca; Rodrigue, James R; Segev, Dorry L

    2015-09-01

    The Best Practice in Live Kidney Donation Consensus Conference held in June of 2014 included the Best Practices in Living Donor Education Workgroup, whose charge was to identify best practice strategies in education of living donors, community outreach initiatives, commercial media, solicitation, and state registries. The workgroup's goal was to identify critical content to include in living kidney donor education and best methods to deliver educational content. A detailed summary of considerations regarding educational content issues for potential living kidney donors is presented, including the consensus that was reached. Educational topics that may require updating on the basis of emerging studies on living kidney donor health outcomes are also presented. Enhancing the educational process is important for increasing living donor comprehension to optimize informed decision-making.

  7. Living Donor Kidney Transplantation: Facilitating Education about Live Kidney Donation--Recommendations from a Consensus Conference.

    PubMed

    Tan, Jane C; Gordon, Elisa J; Dew, Mary Amanda; LaPointe Rudow, Dianne; Steiner, Robert W; Woodle, E Steve; Hays, Rebecca; Rodrigue, James R; Segev, Dorry L

    2015-09-01

    The Best Practice in Live Kidney Donation Consensus Conference held in June of 2014 included the Best Practices in Living Donor Education Workgroup, whose charge was to identify best practice strategies in education of living donors, community outreach initiatives, commercial media, solicitation, and state registries. The workgroup's goal was to identify critical content to include in living kidney donor education and best methods to deliver educational content. A detailed summary of considerations regarding educational content issues for potential living kidney donors is presented, including the consensus that was reached. Educational topics that may require updating on the basis of emerging studies on living kidney donor health outcomes are also presented. Enhancing the educational process is important for increasing living donor comprehension to optimize informed decision-making. PMID:25908792

  8. Who's in the business of saving lives?

    PubMed

    Lee Chang, Pepe

    2006-10-01

    There are individuals, including children, dying needlessly in poverty-stricken third world countries. Many of these deaths could be prevented if pharmaceutical companies provided the drugs needed to save their lives. Some believe that because pharmaceutical companies have the power to save lives, and because they can do so with little effort, they have a special obligation. I argue that there is no distinction, with respect to obligations and responsibilities, between pharmaceutical companies and other types of companies. As a result, to hold pharmaceutical companies especially responsible for saving lives in third world countries is unjustified.

  9. Care services ecosystem for ambient assisted living

    NASA Astrophysics Data System (ADS)

    Camarinha-Matos, Luis M.; Rosas, Joao; Ines Oliveira, Ana; Ferrada, Filipa

    2015-08-01

    Effective provision of care and assistance services in ambient assisted living requires the involvement and collaboration of multiple stakeholders. To support such collaboration, the development of an ecosystem of products and services for active ageing plays an important role. This article introduces a conceptual architecture that supports such care ecosystem. In order to facilitate understanding and better interrelate concepts, a 3-layered model is adopted: Infrastructure layer, Care and assistance services layer and Ambient Assisted Living ecosystem layer. A holistic perspective of ambient assisted living, namely considering four important life settings is adopted: (1) independent living; (2) health and care in life; (3) occupation in life and (4) recreation in life. The proposed architecture is designed in the context of a national Portuguese project and in accordance with the findings of a large European road mapping initiative on ICT and ageing.

  10. Itchy, Scaly Skin? Living with Psoriasis

    MedlinePlus

    ... exit disclaimer . Subscribe Itchy, Scaly Skin? Living With Psoriasis The thick, red, scaly skin of psoriasis can ... Diet Itchy, Scaly Skin? Wise Choices Links Treating Psoriasis Doctors often use a trial-and-error approach ...

  11. Storing data encoded DNA in living organisms

    DOEpatents

    Wong; Pak C. , Wong; Kwong K. , Foote; Harlan P.

    2006-06-06

    Current technologies allow the generation of artificial DNA molecules and/or the ability to alter the DNA sequences of existing DNA molecules. With a careful coding scheme and arrangement, it is possible to encode important information as an artificial DNA strand and store it in a living host safely and permanently. This inventive technology can be used to identify origins and protect R&D investments. It can also be used in environmental research to track generations of organisms and observe the ecological impact of pollutants. Today, there are microorganisms that can survive under extreme conditions. As well, it is advantageous to consider multicellular organisms as hosts for stored information. These living organisms can provide as memory housing and protection for stored data or information. The present invention provides well for data storage in a living organism wherein at least one DNA sequence is encoded to represent data and incorporated into a living organism.

  12. Living as a Breast Cancer Survivor

    MedlinePlus

    ... Emotional aspects of breast cancer Living as a breast cancer survivor For many women with breast cancer, treatment ... making some new choices. Follow-up care after breast cancer treatment Even after you have completed breast cancer ...

  13. Space Station Live: EarthKAM

    NASA Video Gallery

    Space Station Live commentator Pat Ryan interviews Brion Au, EarthKAM Payload Developer. The NASA education program enables middle school students to take pictures of the Earth from the Internation...

  14. A Live Specimen Cell for the Microscope.

    ERIC Educational Resources Information Center

    McNeil, D. W.

    1991-01-01

    Provides background and instructions for the assembly of a microaquarium, or specimen cell, in which the dynamic world of living microorganisms can be viewed through a microscope overextended periods of time utilizing the simplest of materials in the process. (JJK)

  15. Nursing in the sandbox: a lived experience.

    PubMed

    Haynes-Smith, Glennena

    2010-01-01

    Despite an extensive career in military nursing, the author's deployment to Iraq included experiences for which no amount of education could have prepared her: the extreme living conditions, harrowing battlefield injuries, and ethical dilemmas she confronted.

  16. In retrospect: Sixty years of living polymers

    NASA Astrophysics Data System (ADS)

    Patterson, Gary

    2016-08-01

    In the 1950s, the discovery of a class of 'living' polymerization reaction revolutionized the field of polymer science by providing a way of controlling the molecular-weight distribution of polymers. The effects reverberate to this day.

  17. Living with Alpha-1 Antitrypsin Deficiency

    MedlinePlus

    ... Support Living with AAT deficiency may cause fear, anxiety, depression, and stress. Talk about how you feel ... and friends also can help relieve stress and anxiety. Let your loved ones know how you feel ...

  18. Primary students' conceptions of living things

    NASA Astrophysics Data System (ADS)

    Legaspi, Britt Anne

    Elementary school teachers are pressed for time throughout the instructional day to teach all curricular areas as expected by states and districts because of the current focus on reading and mathematics. Thus, foundational science concepts may be overlooked. For example, students' understandings of living and nonliving things may be overlooked by teachers, yet is useful in understanding the nature of living things. In this qualitative study, K-3 grade students were asked to sort objects as either living or nonliving and to give rationales for their choices. It was found that K-3 students readily used physical characteristics, such as having body parts, and physical abilities, such as being able to move, as criteria for living things. Students in grades 1 through 3 were able to articulate their reasons with more adult-like logic based on Jean Piaget' s research on developmental stages.

  19. Healthy Family 2009: Practicing Healthy Adult Living

    MedlinePlus

    ... Navigation Bar Home Current Issue Past Issues Healthy Family 2009 Practicing Healthy Adult Living Past Issues / Winter ... diabetes, or if heart disease runs in your family, begin checking cholesterol at age 20. Colorectal Cancer : ...

  20. Visualizing influenza virus infection in living mice

    PubMed Central

    Pan, Weiqi; Dong, Zhenyuan; Li, Feng; Meng, Weixu; Feng, Liqiang; Niu, Xuefeng; Li, Chufang; Luo, Qinfang; Li, Zhengfeng; Sun, Caijun; Chen, Ling

    2013-01-01

    Preventing and treating influenza virus infection remain a challenge because of incomplete understanding of the host–pathogen interactions, limited therapeutics and lack of a universal vaccine. So far, methods for monitoring the course of infection with influenza virus in real time in living animals are lacking. Here we report the visualization of influenza viral infection in living mice using an engineered replication-competent influenza A virus carrying luciferase reporter gene. After intranasal inoculation, bioluminescence can be detected in the chest and nasopharyngeal passage of living mice. The intensity of bioluminescence in the chest correlates with the dosage of infection and the viral load in the lung. Bioluminescence in the chest of infected mice diminishes on antiviral treatment. This work provides a novel approach that enables real-time study of influenza virus infection and effects of antiviral therapeutics in living animals. PMID:24022374

  1. NES Live Video Chat: Dr. Tara Ruttley

    NASA Video Gallery

    The NASA Extreme Environment Mission Operations program sends groups of NASA employees and contractors to live in Aquarius, an underwater laboratory, for up to three weeks at a time. NEEMO crew mem...

  2. Child Abuse May Shorten Some Women's Lives

    MedlinePlus

    ... page: https://medlineplus.gov/news/fullstory_160478.html Child Abuse May Shorten Some Women's Lives Extreme stress ... 300 middle-aged U.S. adults, female survivors of child abuse were more likely to die over the ...

  3. How Live Performance Moves the Human Heart.

    PubMed

    Shoda, Haruka; Adachi, Mayumi; Umeda, Tomohiro

    2016-01-01

    We investigated how the audience member's physiological reactions differ as a function of listening context (i.e., live versus recorded music contexts). Thirty-seven audience members were assigned to one of seven pianists' performances and listened to his/her live performances of six pieces (fast and slow pieces by Bach, Schumann, and Debussy). Approximately 10 weeks after the live performance, each of the audience members returned to the same room and listened to the recorded performances of the same pianists' via speakers. We recorded the audience members' electrocardiograms in listening to the performances in both conditions, and analyzed their heart rates and the spectral features of the heart-rate variability (i.e., HF/TF, LF/HF). Results showed that the audience's heart rate was higher for the faster than the slower piece only in the live condition. As compared with the recorded condition, the audience's sympathovagal balance (LF/HF) was less while their vagal nervous system (HF/TF) was activated more in the live condition, which appears to suggest that sharing the ongoing musical moments with the pianist reduces the audience's physiological stress. The results are discussed in terms of the audience's superior attention and temporal entrainment to live performance.

  4. How Live Performance Moves the Human Heart

    PubMed Central

    Shoda, Haruka; Adachi, Mayumi; Umeda, Tomohiro

    2016-01-01

    We investigated how the audience member’s physiological reactions differ as a function of listening context (i.e., live versus recorded music contexts). Thirty-seven audience members were assigned to one of seven pianists’ performances and listened to his/her live performances of six pieces (fast and slow pieces by Bach, Schumann, and Debussy). Approximately 10 weeks after the live performance, each of the audience members returned to the same room and listened to the recorded performances of the same pianists’ via speakers. We recorded the audience members’ electrocardiograms in listening to the performances in both conditions, and analyzed their heart rates and the spectral features of the heart-rate variability (i.e., HF/TF, LF/HF). Results showed that the audience’s heart rate was higher for the faster than the slower piece only in the live condition. As compared with the recorded condition, the audience’s sympathovagal balance (LF/HF) was less while their vagal nervous system (HF/TF) was activated more in the live condition, which appears to suggest that sharing the ongoing musical moments with the pianist reduces the audience’s physiological stress. The results are discussed in terms of the audience’s superior attention and temporal entrainment to live performance. PMID:27104377

  5. Just love in live organ donation.

    PubMed

    Zeiler, Kristin

    2009-08-01

    Emotionally-related live organ donation is different from almost all other medical treatments in that a family member or, in some countries, a friend contributes with an organ or parts of an organ to the recipient. Furthermore, there is a long-acknowledged but not well-understood gender-imbalance in emotionally-related live kidney donation. This article argues for the benefit of the concept of just love as an analytic tool in the analysis of emotionally-related live organ donation where the potential donor(s) and the recipient are engaged in a love relation. The concept of just love is helpful in the analysis of these live organ donations even if no statistical gender-imbalance prevails. It is particularly helpful, however, in the analysis of the gender-imbalance in live kidney donations if these donations are seen as a specific kind of care-work, if care-work is experienced as a labour one should perform out of love and if women still experience stronger pressures to engage in care-work than do men. The aim of the article is to present arguments for the need of just love as an analytic tool in the analysis of emotionally-related live organ donation where the potential donor(s) and the recipient are engaged in a love relation. The aim is also to elaborate two criteria that need to be met in order for love to qualify as just and to highlight certain clinical implications.

  6. Educating Prospective Kidney Transplant Recipients and Living Donors about Living Donation: Practical and Theoretical Recommendations for Increasing Living Donation Rates

    PubMed Central

    Waterman, Amy D.; Robbins, Mark L.; Peipert, John D.

    2016-01-01

    A promising strategy for increasing living donor kidney transplant (LDKT) rates is improving education about living donation for both prospective kidney transplant recipients and living donors to help overcome the proven knowledge, psychological, and socioeconomic barriers to LDKT. A recent Consensus Conference on Best Practices in Live Kidney Donation recommended that comprehensive LDKT education be made available to patients at all stages of chronic kidney disease (CKD). However, in considering how to implement this recommendation across different healthcare learning environments, the current lack of available guidance regarding how to design, deliver, and measure the efficacy of LDKT education programs is notable. In the current article, we provide an overview of how one behavior change theory, the Transtheoretical Model of Behavior Change, can guide the delivery of LDKT education for patients at various stages of CKD and readiness for LDKT. We also discuss the importance of creating educational programs for both potential kidney transplant recipients and living donors, and identify key priorities for educational research to reduce racial disparities in LDKT and increase LDKT rates. PMID:27347475

  7. Scanning Ion Conductance Microscopy of Live Keratinocytes

    NASA Astrophysics Data System (ADS)

    Hegde, V.; Mason, A.; Saliev, T.; Smith, F. J. D.; McLean, W. H. I.; Campbell, P. A.

    2012-07-01

    Scanning ion conductance microscopy (SICM) is perhaps the least well known technique from the scanning probe microscopy (SPM) family of instruments. As with its more familiar counterpart, atomic force microscopy (AFM), the technique provides high-resolution topographic imaging, with the caveat that target structures must be immersed in a conducting solution so that a controllable ion current may be utilised as the basis for feedback. In operation, this non-contact characteristic of SICM makes it ideal for the study of delicate structures, such as live cells. Moreover, the intrinsic architecture of the instrument, incorporating as it does, a scanned micropipette, lends itself to combination approaches with complementary techniques such as patch-clamp electrophysiology: SICM therefore boasts the capability for both structural and functional imaging. For the present observations, an ICnano S system (Ionscope Ltd., Melbourn, UK) operating in 'hopping mode' was used, with the objective of assessing the instrument's utility for imaging live keratinocytes under physiological buffers. In scans employing cultured HaCaT cells (spontaneously immortalised, human keratinocytes), we compared the qualitative differences of live cells imaged with SICM and AFM, and also with their respective counterparts after chemical fixation in 4% paraformaldehyde. Characteristic surface microvilli were particularly prominent in live cell imaging by SICM. Moreover, time lapse SICM imaging on live cells revealed that changes in the pattern of microvilli could be tracked over time. By comparison, AFM imaging on live cells, even at very low contact forces (

  8. Amino Acid Substitutions in Cold-Adapted Proteins from Halorubrum lacusprofundi, an Extremely Halophilic Microbe from Antarctica

    PubMed Central

    DasSarma, Shiladitya; Capes, Melinda D.; Karan, Ram; DasSarma, Priya

    2013-01-01

    The halophilic Archaeon Halorubrum lacusprofundi, isolated from the perennially cold and hypersaline Deep Lake in Antarctica, was recently sequenced and compared to 12 Haloarchaea from temperate climates by comparative genomics. Amino acid substitutions for 604 H. lacusprofundi proteins belonging to conserved haloarchaeal orthologous groups (cHOGs) were determined and found to occur at 7.85% of positions invariant in proteins from mesophilic Haloarchaea. The following substitutions were observed most frequently: (a) glutamic acid with aspartic acid or alanine; (b) small polar residues with other small polar or non-polar amino acids; (c) small non-polar residues with other small non-polar residues; (d) aromatic residues, especially tryptophan, with other aromatic residues; and (e) some larger polar residues with other similar residues. Amino acid substitutions for a cold-active H. lacusprofundi β-galactosidase were then examined in the context of a homology modeled structure at residues invariant in homologous enzymes from mesophilic Haloarchaea. Similar substitutions were observed as in the genome-wide approach, with the surface accessible regions of β-galactosidase displaying reduced acidity and increased hydrophobicity, and internal regions displaying mainly subtle changes among smaller non-polar and polar residues. These findings are consistent with H. lacusprofundi proteins displaying amino acid substitutions that increase structural flexibility and protein function at low temperature. We discuss the likely mechanisms of protein adaptation to a cold, hypersaline environment on Earth, with possible relevance to life elsewhere. PMID:23536799

  9. A cold-adapted esterase of a novel marine isolate, Pseudoalteromonas arctica: gene cloning, enzyme purification and characterization.

    PubMed

    Al Khudary, Rami; Venkatachalam, Ramprasath; Katzer, Moritz; Elleuche, Skander; Antranikian, Garabed

    2010-05-01

    A gene encoding an esterase (estO) was identified and sequenced from a gene library screen of the psychrotolerant bacterium Pseudoalteromonas arctica. Analysis of the 1,203 bp coding region revealed that the deduced peptide sequence is composed of 400 amino acids with a predicted molecular mass of 44.1 kDa. EstO contains a N-terminal esterase domain and an additional OsmC domain at the C-terminus (osmotically induced family of proteins). The highly conserved five-residue motif typical for all alpha/beta hydrolases (G x S x G) was detected from position 104 to 108 together with a putative catalytic triad consisting of Ser(106), Asp(196), and His(225). Sequence comparison showed that EstO exhibits 90% amino acid identity with hypothetical proteins containing similar esterase and OsmC domains but only around 10% identity to the amino acid sequences of known esterases. EstO variants with and without the OsmC domain were produced and purified as His-tag fusion proteins in E. coli. EstO displayed an optimum pH of 7.5 and optimum temperature of 25 degrees C with more than 50% retained activity at the freezing point of water. The thermostability of EstO (50% activity after 5 h at 40 degrees C) dramatically increased in the truncated variant (50% activity after 2.5 h at 90 degrees C). Furthermore, the esterase displays broad substrate specificity for esters of short-chain fatty acids (C(2)-C(8)).

  10. Expression and enzymatic characterization of a cold-adapted β-agarase from Antarctic bacterium Pseudoalteromonas sp. NJ21

    NASA Astrophysics Data System (ADS)

    Li, Jiang; Sha, Yujie

    2015-03-01

    An agar-degrading bacterium, designated as Pseudoalteromonas sp. NJ21, was isolated from an Antarctic sediment sample. The agarase gene aga1161 from Pseudoalteromonas sp. NJ21 consisting of a 2 382-bp coding region was cloned. The gene encodes a 793-amino acids protein and was found to possess characteristic features of the Glyco_hydro_42 family. The recombinant agarase (rAga1161) was overexpressed in Escherichia coli and purified as a fusion protein. Enzyme activity analysis revealed that the optimum temperature and pH for the purified recombinant agarase were 30-40°C and 8.0, respectively. rAga1161 was found to maintain as much as 80% of its maximum activity at 10°C, which is typical of a coldadapted enzyme. The pattern of agar hydrolysis demonstrated that the enzyme is an β-agarase, producing neoagarobiose (NA2) as the final main product. Furthermore, this work is the first proof of an agarolytic activity in Antarctic bacteria and these results indicate the potential for the Antarctic agarase as a catalyst in medicine, food and cosmetic industries.

  11. Translation during cold adaptation does not involve mRNA-rRNA base pairing through the downstream box.

    PubMed

    La Teana, A; Brandi, A; O'Connor, M; Freddi, S; Pon, C L

    2000-10-01

    The downstream box (DB) has been proposed to enhance translation of several mRNAs and to be a key element controlling the expression of cold-shocked mRNAs. However, the proposal that the DB exerts its effects through a base pairing interaction with the complementary anti-downstream box (antiDB) sequence (nt 1469-1483) located in the penultimate stem (helix 44) of 16S rRNA remains controversial. The existence of this interaction during initiation of protein synthesis under cold-shock conditions has been investigated in the present work using an Escherichia coli strain whose ribosomes lack the potential to base pair with mRNA because of a 12 bp inversion of the antiDB sequence in helix 44. Our results show that this strain is capable of cold acclimation, withstands cold shock, and its ribosomes translate mRNAs that contain or lack DB sequences with similar efficiency, comparable to that of the wild type. The structure of helix 44 in 30S ribosomal subunits from cells grown at 37 degrees C and from cells subjected to cold shock was also analyzed by binding a 32P-labeled oligonucleotide complementary to the antiDB region and by chemical probing with DMS and kethoxal. Both approaches clearly indicate that this region is in a double-stranded conformation and therefore not available for base pairing with mRNA.

  12. Characterization of an acidic cold-adapted cutinase from Thielavia terrestris and its application in flavor ester synthesis.

    PubMed

    Xu, Haibo; Yan, Qiaojuan; Duan, Xiaojie; Yang, Shaoqing; Jiang, Zhengqiang

    2015-12-01

    An acidic cutinase (TtcutB) from Thielavia terrestris CAU709 was purified to apparent homogeneity with 983 Um g(-1) specific activity. The molecular mass of the enzyme was estimated to be 27.3 and 27.9 kDa by SDS-PAGE and gel filtration, respectively. A peptide sequence homology search revealed no homologous cutinases from T. terrestris, except for one putative cutinase gene (XP003656017.1), indicating that TtcutB is a novel enzyme. TtcutB exhibited an acidic pH optimum of 4.0, and stability at pH 2.5-10.5. Optimal activity was at 55 °C, it was stable up to 65 °C, and retained over 30% activity at 0 °C. Km values toward p-nitrophenyl (pNP) acetate, pNP-butyrate and pNP-caproate were 8.3, 1.1 and 0.88 mM, respectively. The cutinase exhibited strong synthetic activity on flavor ester butyl butyrate under non-aqueous environment, and the highest esterification efficiency of 95% was observed under the optimized reaction conditions. The enzyme's unique biochemical properties suggest great potential in flavor esters-producing industries.

  13. Diversity and cold adaptation of culturable endophytic fungi from bryophytes in the Fildes Region, King George Island, maritime Antarctica.

    PubMed

    Zhang, Tao; Zhang, Yu-Qin; Liu, Hong-Yu; Wei, Yu-Zhen; Li, Hai-Long; Su, Jing; Zhao, Li-Xun; Yu, Li-Yan

    2013-04-01

    Endophytic fungi associated with three bryophyte species in the Fildes Region, King George Island, maritime Antarctica, that is, the liverwort Barbilophozia hatcheri, the mosses Chorisodontium aciphyllum and Sanionia uncinata, were studied by culture-dependent method. A total of 128 endophytic fungi were isolated from 1329 tissue segments of 14 samples. The colonization rate of endophytic fungi in three bryophytes species were 12.3%, 12.1%, and 8.7%, respectively. These isolates were identified to 21 taxa, with 15 Ascomycota, 5 Basidiomycota, and 1 unidentified fungus, based on morphological characteristics and sequence analyses of ITS region and D1/D2 domain. The dominant fungal endophyte was Hyaloscyphaceae sp. in B. hatcheri, Rhizoscyphus sp. in C. aciphyllum, and one unidentified fungus in S. uncinata; and their relative frequencies were 33.3%, 32.1%, and 80.0%, respectively. Furthermore, different Shannon-Weiner diversity indices (0.91-1.99) for endophytic fungi and low endophytic fungal composition similarities (0.19-0.40) were found in three bryophyte species. Growth temperature tests indicated that 21 taxa belong to psychrophiles (9), psychrotrophs (11), and mesophile (1). The results herein demonstrate that the Antarctic bryophytes are an interesting source of fungal endophytes and the endophytic fungal composition is different among the bryophyte species, and suggest that these fungal endophytes are adapted to cold stress in Antarctica.

  14. Isolation and Characterization of a Novel Cold-Adapted Esterase, MtEst45, from Microbulbifer thermotolerans DAU221

    PubMed Central

    Lee, Yong-Suk

    2016-01-01

    A novel esterase, MtEst45, was isolated from a fosmid genomic library of Microbulbifer thermotolerans DAU221. The encoding gene is predicted to have a mass of 45,564 Da and encodes 495 amino acids, excluding a 21 amino acid signal peptide. MtEst45 showed a low amino acid identity (approximately 23–24%) compared with other lipolytic enzymes belonging to Family III, a closely related bacterial lipolytic enzyme family. MtEst45 also showed a conserved GXSXG motif, G131IS133YG135, which was reported as active site of known lipolytic enzymes, and the putative catalytic triad composed of D237 and H265. Because these mutants of MtEst45, which was S133A, D237N, and H265L, had no activity, these catalytic triad is deemed essential for the enzyme catalysis. MtEst45 was overexpressed in Escherichia coli BL21 (DE3) and purified via His-tag affinity chromatography. The optimal pH and temperature of MtEst45 were estimated to be 8.17 and 46.27°C by response surface methodology, respectively. Additionally, MtEst45 was also active between 1 and 15°C. The optimal hydrolysis substrate for MtEst45 among p-nitrophenyl esters (C2–C18) was p-nitrophenyl butyrate, and the Km and Vmax values were 0.0998 mM and 550 μmol/min/mg of protein, respectively. MtEst45 was strongly inhibited by Hg2+, Zn2+, and Cu2+ ions; by phenylmethanesulfonyl fluoride; and by β-mercaptoethanol. Ca2+ did not affect the enzyme's activity. These biochemical properties, sequence identity, and phylogenetic analysis suggest that MtEst45 represents a novel and valuable bacterial lipolytic enzyme family and is useful for biotechnological applications. PMID:26973604

  15. Cold-adapted digestive aspartic protease of the clawed lobsters Homarus americanus and Homarus gammarus: biochemical characterization.

    PubMed

    Rojo, Liliana; García-Carreño, Fernando; de Los Angeles Navarrete del Toro, Maria

    2013-02-01

    Aspartic proteinases in the gastric fluid of clawed lobsters Homarus americanus and Homarus gammarus were isolated to homogeneity by single-step pepstatin-A affinity chromatography; such enzymes have been previously identified as cathepsin D-like enzymes based on their deduced amino acid sequence. Here, we describe their biochemical characteristics; the properties of the lobster enzymes were compared with those of its homolog, bovine cathepsin D, and found to be unique in a number of ways. The lobster enzymes demonstrated hydrolytic activity against synthetic and natural substrates at a wider range of pH; they were more temperature-sensitive, showed no changes in the K(M) value at 4°C, 10°C, and 25°C, and had 20-fold higher k(cat)/K(M) values than bovine enzyme. The bovine enzyme was temperature-dependent. We propose that both properties arose from an increase in molecular flexibility required to compensate for the reduction of reaction rates at low habitat temperatures. This is supported by the fast denaturation rates induced by temperature. PMID:22648335

  16. Stereochemical course of hydrolytic reaction catalyzed by alpha-galactosidase from cold adaptable marine bacterium of genus Pseudoalteromonas

    NASA Astrophysics Data System (ADS)

    Bakunina, Irina; Balabanova, Larissa; Golotin, Vasiliy; Slepchenko, Lyubov; Isakov, Vladimir; Rasskazov, Valeriy

    2014-10-01

    The recombinant α-galactosidase of the marine bacterium (α-PsGal) was synthesized with the use of the plasmid 40Gal, consisting of plasmid pET-40b (+) (Novagen) and the gene corresponding to the open reading frame of the mature α-galactosidase of marine bacterium Pseudoalteromonas sp. KMM 701, transformed into the E. coli Rosetta(DE3) cells. In order to understand the mechanism of action, the stereochemistry of hydrolysis of 4-nitrophenyl α-D-galactopyranoside (4-NPGP) by α-PsGal was measured by 1H NMR spectroscopy. The kinetics of formation of α- and β-anomer of galactose showed that α-anomer initially formed and accumulated, and then an appreciable amount of β-anomer appeared as a result of mutarotation. The data clearly show that the enzymatic hydrolysis of 4-NPGP proceeds with the retention of anomeric configuration, probably, due to a double displacement mechanism of reaction.

  17. A novel cold-adapted β-galactosidase isolated from Halomonas sp. S62: gene cloning, purification and enzymatic characterization.

    PubMed

    Wang, Guo-Xiang; Gao, Yun; Hu, Bo; Lu, Xiao-Ling; Liu, Xiao-Yu; Jiao, Bing-Hua

    2013-08-01

    A novel 1,170 bp β-galactosidase gene sequence from Halomonas sp. S62 (BGalH) was identified through whole genome sequencing and was submitted to GenBank (Accession No. JQ337961). The BGalH gene was heterologously expressed in Escherichia coli BL21(DE3) cells, and the enzymatic properties of recombinant BGalH were studied. According to the polyacrylamide gel electrophoresis results and the sequence alignment analysis, BGalH is a dimeric protein and cannot be classified into one of the known β-galactosidase families (GH1, GH2, GH35, GH42). The optimal pH and temperature were determined to be 7.0 and 45 °C, respectively; the K m and K cat were 2.9 mM and 390.3 s(-1), respectively, for the reaction with the substrate ortho-nitrophenyl-β-D-galactopyranoside. At 0-20 °C, BGalH exhibited 50-70 % activity relative to its activity under the optimal conditions. BGalH was stable over a wide range of pHs (6.0-8.5) after a 1 h incubation (>93 % relative activity) and was thermostable at 50 °C and below (>60 % relative activity). The enzyme hydrolyzes lactose completely in milk over 24 h at 7 °C. The characteristics of this novel β-galactosidase suggest that BGalH may be a good candidate for medical researches and food industry applications.

  18. Diversification of the cold-adapted butterfly genus Oeneis related to Holarctic biogeography and climatic niche shifts.

    PubMed

    Kleckova, I; Cesanek, M; Fric, Z; Pellissier, L

    2015-11-01

    Both geographical and ecological speciation interact during the evolution of a clade, but the relative contribution of these processes is rarely assessed for cold-dwelling biota. Here, we investigate the role of biogeography and the evolution of ecological traits on the diversification of the Holarctic arcto-alpine butterfly genus Oeneis (Lepidoptera: Satyrinae). We reconstructed the molecular phylogeny of the genus based on one mitochondrial (COI) and three nuclear (GAPDH, RpS5, wingless) genes. We inferred the biogeographical scenario and the ancestral state reconstructions of climatic and habitat requirements. Within the genus, we detected five main species groups corresponding to the taxonomic division and further paraphyletic position of Neominois (syn. n.). Next, we transferred O. aktashi from the hora to the polixenes species group on the bases of molecular relationships. We found that the genus originated in the dry grasslands of the mountains of Central Asia and dispersed over the Beringian Land Bridges to North America several times independently. Holarctic mountains, in particular the Asian Altai Mts. and Sayan Mts., host the oldest lineages and most of the species diversity. Arctic species are more recent, with Pliocene or Pleistocene origin. We detected a strong phylogenetic signal for the climatic niche, where one lineage diversified towards colder conditions. Altogether, our results indicate that both dispersal across geographical areas and occupation of distinct climatic niches promoted the diversification of the Oeneis genus. PMID:26166775

  19. Crystal structure analysis of a glycosides hydrolase family 42 cold-adapted ß-galactosidase from Rahnella sp. R3

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The ß-galactosidase isolated from a psychrotrophic bacterium, Rahnella sp. R3 (R-ß-Gal), exhibits high activity at low temperature. R-ß-Gal is a member of the glycoside hydrolases family 42 (GH42), and forms a 225 kDa trimeric structure in solution. The X-ray crystal structure of R-ß-Gal was determi...

  20. Microorganisms resistant to free-living amoebae.

    PubMed

    Greub, Gilbert; Raoult, Didier

    2004-04-01

    Free-living amoebae feed on bacteria, fungi, and algae. However, some microorganisms have evolved to become resistant to these protists. These amoeba-resistant microorganisms include established pathogens, such as Cryptococcus neoformans, Legionella spp., Chlamydophila pneumoniae, Mycobacterium avium, Listeria monocytogenes, Pseudomonas aeruginosa, and Francisella tularensis, and emerging pathogens, such as Bosea spp., Simkania negevensis, Parachlamydia acanthamoebae, and Legionella-like amoebal pathogens. Some of these amoeba-resistant bacteria (ARB) are lytic for their amoebal host, while others are considered endosymbionts, since a stable host-parasite ratio is maintained. Free-living amoebae represent an important reservoir of ARB and may, while encysted, protect the internalized bacteria from chlorine and other biocides. Free-living amoebae may act as a Trojan horse, bringing hidden ARB within the human "Troy," and may produce vesicles filled with ARB, increasing their transmission potential. Free-living amoebae may also play a role in the selection of virulence traits and in adaptation to survival in macrophages. Thus, intra-amoebal growth was found to enhance virulence, and similar mechanisms seem to be implicated in the survival of ARB in response to both amoebae and macrophages. Moreover, free-living amoebae represent a useful tool for the culture of some intracellular bacteria and new bacterial species that might be potential emerging pathogens.

  1. Live ultrasound volume reconstruction using scout scanning

    PubMed Central

    Meyer, Amelie; Lasso, Andras; Ungi, Tamas; Fichtinger, Gabor

    2015-01-01

    INTRODUCTION Ultrasound-guided interventions often necessitate scanning of deep-seated anatomical structures that may be hard to visualize. Visualization can be improved using reconstructed 3D ultrasound volumes. High-resolution 3D reconstruction of a large area during clinical interventions is challenging if the region of interest is unknown. We propose a two-stage scanning method allowing the user to perform quick low-resolution scouting followed by high-resolution live volume reconstruction. METHODS Scout scanning is accomplished by stacking 2D tracked ultrasound images into a low-resolution volume. Then, within a region of interest defined in the scout scan, live volume reconstruction can be performed by continuous scanning until sufficient image density is achieved. We implemented the workflow as a module of the open-source 3D Slicer application, within the SlicerIGT extension and building on the PLUS toolkit. RESULTS Scout scanning is performed in a few seconds using 3 mm spacing to allow region of interest definition. Live reconstruction parameters are set to provide good image quality (0.5 mm spacing, hole filling enabled) and feedback is given during live scanning by regularly updated display of the reconstructed volume. DISCUSSION Use of scout scanning may allow the physician to identify anatomical structures. Subsequent live volume reconstruction in a region of interest may assist in procedures such as targeting needle interventions or estimating brain shift during surgery. PMID:26005249

  2. Microorganisms Resistant to Free-Living Amoebae

    PubMed Central

    Greub, Gilbert; Raoult, Didier

    2004-01-01

    Free-living amoebae feed on bacteria, fungi, and algae. However, some microorganisms have evolved to become resistant to these protists. These amoeba-resistant microorganisms include established pathogens, such as Cryptococcus neoformans, Legionella spp., Chlamydophila pneumoniae, Mycobacterium avium, Listeria monocytogenes, Pseudomonas aeruginosa, and Francisella tularensis, and emerging pathogens, such as Bosea spp., Simkania negevensis, Parachlamydia acanthamoebae, and Legionella-like amoebal pathogens. Some of these amoeba-resistant bacteria (ARB) are lytic for their amoebal host, while others are considered endosymbionts, since a stable host-parasite ratio is maintained. Free-living amoebae represent an important reservoir of ARB and may, while encysted, protect the internalized bacteria from chlorine and other biocides. Free-living amoebae may act as a Trojan horse, bringing hidden ARB within the human “Troy,” and may produce vesicles filled with ARB, increasing their transmission potential. Free-living amoebae may also play a role in the selection of virulence traits and in adaptation to survival in macrophages. Thus, intra-amoebal growth was found to enhance virulence, and similar mechanisms seem to be implicated in the survival of ARB in response to both amoebae and macrophages. Moreover, free-living amoebae represent a useful tool for the culture of some intracellular bacteria and new bacterial species that might be potential emerging pathogens. PMID:15084508

  3. Live ultrasound volume reconstruction using scout scanning

    NASA Astrophysics Data System (ADS)

    Meyer, Amelie; Lasso, Andras; Ungi, Tamas; Fichtinger, Gabor

    2015-03-01

    Ultrasound-guided interventions often necessitate scanning of deep-seated anatomical structures that may be hard to visualize. Visualization can be improved using reconstructed 3D ultrasound volumes. High-resolution 3D reconstruction of a large area during clinical interventions is challenging if the region of interest is unknown. We propose a two-stage scanning method allowing the user to perform quick low-resolution scouting followed by high-resolution live volume reconstruction. Scout scanning is accomplished by stacking 2D tracked ultrasound images into a low-resolution volume. Then, within a region of interest defined in the scout scan, live volume reconstruction can be performed by continuous scanning until sufficient image density is achieved. We implemented the workflow as a module of the open-source 3D Slicer application, within the SlicerIGT extension and building on the PLUS toolkit. Scout scanning is performed in a few seconds using 3 mm spacing to allow region of interest definition. Live reconstruction parameters are set to provide good image quality (0.5 mm spacing, hole filling enabled) and feedback is given during live scanning by regularly updated display of the reconstructed volume. Use of scout scanning may allow the physician to identify anatomical structures. Subsequent live volume reconstruction in a region of interest may assist in procedures such as targeting needle interventions or estimating brain shift during surgery.

  4. [Living donor liver transplantation in adults].

    PubMed

    Neumann, U P; Neuhaus, P; Schmeding, M

    2010-09-01

    The worldwide shortage of adequate donor organs implies that living donor liver transplantation represents a valuable alternative to cadaveric transplantation. In addition to the complex surgical procedure the correct identification of eligible donors and recipients plays a decisive role in living donor liver transplantation. Donor safety must be of ultimate priority and overrules all other aspects involved. In contrast to the slightly receding numbers in Europe and North America, in recent years Asian programs have enjoyed constantly increasing living donor activity. The experience of the past 15 years has clearly demonstrated that technical challenges of both bile duct anastomosis and venous outflow of the graft significantly influence postoperative outcome. While short-term in-hospital morbidity remains increased compared to cadaveric transplantation, long-term survival of both graft and patient are comparable or even better than in deceased donor transplantation. Especially for patients expecting long waiting times under the MELD allocation system, living donor liver transplantation offers an excellent therapeutic alternative. Expanding the so-called "Milan criteria" for HCC patients with the option for living donor liver transplantation is currently being controversially debated.

  5. Comparison of egg and high yielding MDCK cell-derived live attenuated influenza virus for commercial production of trivalent influenza vaccine: in vitro cell susceptibility and influenza virus replication kinetics in permissive and semi-permissive cells.

    PubMed

    Hussain, Althaf I; Cordeiro, Melissa; Sevilla, Elizabeth; Liu, Jonathan

    2010-05-14

    Currently MedImmune manufactures cold-adapted (ca) live, attenuated influenza vaccine (LAIV) from specific-pathogen free (SPF) chicken eggs. Difficulties in production scale-up and potential exposure of chicken flocks to avian influenza viruses especially in the event of a pandemic influenza outbreak have prompted evaluation and development of alternative non-egg based influenza vaccine manufacturing technologies. As part of MedImmune's effort to develop the live attenuated influenza vaccine (LAIV) using cell culture production technologies we have investigated the use of high yielding, cloned MDCK cells as a substrate for vaccine production by assessing host range and virus replication of influenza virus produced from both SPF egg and MDCK cell production technologies. In addition to cloned MDCK cells the indicator cell lines used to evaluate the impact of producing LAIV in cells on host range and replication included two human cell lines: human lung carcinoma (A549) cells and human muco-epidermoid bronchiolar carcinoma (NCI H292) cells. The influenza viruses used to infect the indicators cell lines represented both the egg and cell culture manufacturing processes and included virus strains that composed the 2006-2007 influenza seasonal trivalent vaccine (A/New Caledonia/20/99 (H1N1), A/Wisconsin/67/05 (H3N2) and B/Malaysia/2506/04). Results from this study demonstrate remarkable similarity between influenza viruses representing the current commercial egg produced and developmental MDCK cell produced vaccine production platforms. MedImmune's high yielding cloned MDCK cells used for the cell culture based vaccine production were highly permissive to both egg and cell produced ca attenuated influenza viruses. Both the A549 and NCI H292 cells regardless of production system were less permissive to influenza A and B viruses than the MDCK cells. Irrespective of the indicator cell line used the replication properties were similar between egg and the cell produced

  6. Lived religion: implications for nursing ethics.

    PubMed

    Reimer-Kirkham, Sheryl

    2009-07-01

    This article explores how ethics and religion interface in everyday life by drawing on a study examining the negotiation of religious and spiritual plurality in health care. Employing methods of critical ethnography, namely, interviews and participant observation, data were collected from patients, health care providers, administrators and spiritual care providers. The findings revealed the degree to which 'lived religion' was intertwined with 'lived ethics' for many participants; particularly for people from the Sikh faith. For these participants, religion was woven into everyday life, making distinctions between public and private, secular and sacred spaces improbable. Individual interactions, institutional resource allocation, and social discourses are all embedded in social relationships of power that prevent religion from being a solely personal or private matter. Strategies for the reintegration of religion into nursing ethics are: adjusting professional codes and theories of ethics to reflect the influence of religion; and the contribution of critical perspectives, such as postcolonial feminism, to the understanding of lived ethics.

  7. Assessment of Live Plague Vaccine Candidates.

    PubMed

    Feodorova, Valentina A; Sayapina, Lidiya V; Motin, Vladimir L

    2016-01-01

    Since its creation in the early twentieth century, live plague vaccine EV has been successfully applied to millions of people without severe complications. This vaccine has been proven to elicit protection against both bubonic and pneumonic plague, and it is still in use in populations at risk mainly in the countries of the former Soviet Union. Despite extensive efforts in developing subunit vaccines, there is a reviving interest in creation of a precisely attenuated strain of Yersinia pestis superior to the EV that can serve as a live plague vaccine with improved characteristics. Here we summarize decades of experience of the Russian anti-plague research in developing a standard protocol for early-stage evaluation of safety and immunogenicity of live plague vaccines. This protocol allows step-by-step comparison of the novel test candidates with the EV vaccine by using subcutaneous immunization and bubonic plague infection models in two animal species, e.g., guinea pigs and mice. PMID:27076149

  8. Role of live yeasts in rumen ecosystem.

    PubMed

    Oeztuerk, Hakan; Sagmanligil, Vedat

    2009-07-01

    For many years, ruminant nutritionists and microbiologists have been interested in manipulating the microbial ecosystem of the rumen to improve production efficiency by domestic ruminants. Antibiotic ionophores have been used successfully for this purpose. However, the use of antibiotics in animal feeds has been banned in the European Union since January 2006 due to the risk of spreading antibiotic resistance. For this reason, scientists have become interested in evaluating other alternatives to control specific microbial populations to modulate rumen fermentation. Dietary supplements of live yeast preparations, based on Saccharomyces cerevisiae, have been reported to improve health and productivity of ruminants. In contrast to antimicrobial agents, live yeasts offer a natural alternative to manipulate animal performance. This review discusses the modes of action of live yeasts in rumen ecosystem and their subsequent effects on animal performance. PMID:19753793

  9. The Vernacular Landscape of Assisted Living

    PubMed Central

    Roth, Erin G.; Eckert, J. Kevin

    2011-01-01

    This article is an exploration into the vernacular landscape of Assisted Living (AL), a conceptual idea borrowed from cultural geographer J.B. Jackson, which distinguishes formalized, planned space from those spaces which are unintended and often created spontaneously--vernacular. Based upon three large-scale, multi-year ethnographic studies in Maryland, we consider some of the ways people who live in AL relate to and respond to the built environment, at times subverting the intended purpose of design to make it their own. The conflict that often ensues over both planned and vernacular public and private space, we propose is ultimately the product of living within an environment that is both someone’s home as well as a place of business, whose job it is to keep people safe. Within this physical context of vernacular private and public spaces, this article enriches understandings about the way autonomy and privacy expresses itself. PMID:21949467

  10. Live-cell imaging of cyanobacteria.

    PubMed

    Yokoo, Rayka; Hood, Rachel D; Savage, David F

    2015-10-01

    Cyanobacteria are a diverse bacterial phylum whose members possess a high degree of ultrastructural organization and unique gene regulatory mechanisms. Unraveling this complexity will require the use of live-cell fluorescence microscopy, but is impeded by the inherent fluorescent background associated with light-harvesting pigments and the need to feed photosynthetic cells light. Here, we outline a roadmap for overcoming these challenges. Specifically, we show that although basic cyanobacterial biology creates challenging experimental constraints, these restrictions can be mitigated by the careful choice of fluorophores and microscope instrumentation. Many of these choices are motivated by recent successful live-cell studies. We therefore also highlight how live-cell imaging has advanced our understanding of bacterial microcompartments, circadian rhythm, and the organization and segregation of the bacterial nucleoid.

  11. The Vernacular Landscape of Assisted Living.

    PubMed

    Roth, Erin G; Eckert, J Kevin

    2011-08-01

    This article is an exploration into the vernacular landscape of Assisted Living (AL), a conceptual idea borrowed from cultural geographer J.B. Jackson, which distinguishes formalized, planned space from those spaces which are unintended and often created spontaneously--vernacular. Based upon three large-scale, multi-year ethnographic studies in Maryland, we consider some of the ways people who live in AL relate to and respond to the built environment, at times subverting the intended purpose of design to make it their own. The conflict that often ensues over both planned and vernacular public and private space, we propose is ultimately the product of living within an environment that is both someone's home as well as a place of business, whose job it is to keep people safe. Within this physical context of vernacular private and public spaces, this article enriches understandings about the way autonomy and privacy expresses itself. PMID:21949467

  12. Nucleic Acid Aptamers for Living Cell Analysis

    NASA Astrophysics Data System (ADS)

    Xiong, Xiangling; Lv, Yifan; Chen, Tao; Zhang, Xiaobing; Wang, Kemin; Tan, Weihong

    2014-06-01

    Cells as the building blocks of life determine the basic functions and properties of a living organism. Understanding the structure and components of a cell aids in the elucidation of its biological functions. Moreover, knowledge of the similarities and differences between diseased and healthy cells is essential to understanding pathological mechanisms, identifying diagnostic markers, and designing therapeutic molecules. However, monitoring the structures and activities of a living cell remains a challenging task in bioanalytical and life science research. To meet the requirements of this task, aptamers, as “chemical antibodies,” have become increasingly powerful tools for cellular analysis. This article reviews recent advances in the development of nucleic acid aptamers in the areas of cell membrane analysis, cell detection and isolation, real-time monitoring of cell secretion, and intracellular delivery and analysis with living cell models. Limitations of aptamers and possible solutions are also discussed.

  13. Bioluminescence imaging in live cells and animals.

    PubMed

    Tung, Jack K; Berglund, Ken; Gutekunst, Claire-Anne; Hochgeschwender, Ute; Gross, Robert E

    2016-04-01

    The use of bioluminescent reporters in neuroscience research continues to grow at a rapid pace as their applications and unique advantages over conventional fluorescent reporters become more appreciated. Here, we describe practical methods and principles for detecting and imaging bioluminescence from live cells and animals. We systematically tested various components of our conventional fluorescence microscope to optimize it for long-term bioluminescence imaging. High-resolution bioluminescence images from live neurons were obtained with our microscope setup, which could be continuously captured for several hours with no signs of phototoxicity. Bioluminescence from the mouse brain was also imaged noninvasively through the intact skull with a conventional luminescence imager. These methods demonstrate how bioluminescence can be routinely detected and measured from live cells and animals in a cost-effective way with common reagents and equipment.

  14. Ecology of free-living metacercariae (Trematoda).

    PubMed

    Morley, Neil J

    2015-06-01

    The presence of trematodes with a free-living metacercarial stage is a common feature of most habitats and includes important species such as Fasciola hepatica, Parorchis acanthus and Zygocotyle lunata. These trematodes encyst on the surface of an animal or plant that can act as a transport host, which form the diet of the target definitive host. Although these species are often considered individually, they display common characteristics in their free-living biology indicating a shared transmission strategy, yet in comparison to species with penetrative cercariae this aspect of their life cycles remains much overlooked. This review integrates the diverse data and presents a novel synthesis of free-living metacercariae using epibiosis as the basis of a new framework to describe the relationship between transport hosts and parasites. All aspects of their biology during the period that they are metabolically independent of a host are considered, from cercarial emergence to metacercarial excystment.

  15. Chlamydial infections in free-living birds

    USGS Publications Warehouse

    Brand, C.J.

    1989-01-01

    Most studies of chlamydial infections in free-living wild birds have been limited to surveys for the presence of Chlamydia psittaci or antibody to C psittaci and have largely been done in association with the identification of chlamydiosis in human beings, commercial fowl, or pet birds. The emphasis of these studies has been to determine the prevalence of infection and the potential role of wild birds in the spread of chlamydiae to domestic birds and human beings. Little is known about the epizootiology of chlamydiosis in free-living birds or its affect on their population dynamics. The following article is a summary of reported studies of chlamydiosis in free-living wild birds in relation to host range, ecologic aspects of transmission and maintenance, and the prevalence of disease.

  16. Assessment of Live Plague Vaccine Candidates.

    PubMed

    Feodorova, Valentina A; Sayapina, Lidiya V; Motin, Vladimir L

    2016-01-01

    Since its creation in the early twentieth century, live plague vaccine EV has been successfully applied to millions of people without severe complications. This vaccine has been proven to elicit protection against both bubonic and pneumonic plague, and it is still in use in populations at risk mainly in the countries of the former Soviet Union. Despite extensive efforts in developing subunit vaccines, there is a reviving interest in creation of a precisely attenuated strain of Yersinia pestis superior to the EV that can serve as a live plague vaccine with improved characteristics. Here we summarize decades of experience of the Russian anti-plague research in developing a standard protocol for early-stage evaluation of safety and immunogenicity of live plague vaccines. This protocol allows step-by-step comparison of the novel test candidates with the EV vaccine by using subcutaneous immunization and bubonic plague infection models in two animal species, e.g., guinea pigs and mice.

  17. Living with rattlesnakes: Chernobyl and Dounreay.

    PubMed

    Roff, S R

    2000-01-01

    After the Three Mile Island nuclear plant accident in 1979, a local Pennsylvania resident told The New York Times 'You live with that plant over there for years and years and don't think much about it. But it's like living with a rattlesnake. Sooner or later it's going to bite you. You just don't know when.' As we enter the twenty-first century, many communities find themselves living with rattlesnakes--one at least of which has already bitten. This article considers the official responses to Chernobyl in the first decade after the accident and compares them to the statements surrounding the sudden 1998 decision to decommission the nuclear reprocessing plant at Dounreay in Scotland. PMID:10893942

  18. Environmental assessment requirements for live biological drugs.

    PubMed

    Sutton, Ann

    2008-02-01

    Marketing approval of biological products by the US Food and Drug Administration must comply with requirements of Code of Federal Regulations title 21 part 25, "Environmental Impact Considerations." An environmental impact statement is usually not required. Environmental assessment is required unless excluded. As naturally occurring substances, biological products qualify for categorical exclusion if manufacture and use do not significantly alter their concentration or distribution in the human environment. The manufacturing process and establishment descriptions in the license application should include enough detail to ensure that waste is controlled and inactivated. During clinical development of a live biotherapeutic product, data should be collected regarding the shedding of live organisms from treated patients. The ability of the live organism to persist in the environment should be assessed, and instructions for safe handling by health care providers and consumers should be incorporated into the package insert.

  19. Lived religion: implications for nursing ethics.

    PubMed

    Reimer-Kirkham, Sheryl

    2009-07-01

    This article explores how ethics and religion interface in everyday life by drawing on a study examining the negotiation of religious and spiritual plurality in health care. Employing methods of critical ethnography, namely, interviews and participant observation, data were collected from patients, health care providers, administrators and spiritual care providers. The findings revealed the degree to which 'lived religion' was intertwined with 'lived ethics' for many participants; particularly for people from the Sikh faith. For these participants, religion was woven into everyday life, making distinctions between public and private, secular and sacred spaces improbable. Individual interactions, institutional resource allocation, and social discourses are all embedded in social relationships of power that prevent religion from being a solely personal or private matter. Strategies for the reintegration of religion into nursing ethics are: adjusting professional codes and theories of ethics to reflect the influence of religion; and the contribution of critical perspectives, such as postcolonial feminism, to the understanding of lived ethics. PMID:19528098

  20. Effect of bait in live trapping Peromyscus

    USGS Publications Warehouse

    Stickel, L.F.

    1948-01-01

    SUMMARY: Evidence from live trapping tests indicated that Peromyscus leucopus did not leave their home ranges because of the attraction of trap bait in nearby areas. A trap line down the center of a heavily live-trapped area caught as many mice before the area trapping as afterward. Thus, there was reason to believe that the area trapping did not serve to pre-bait the mice. Two unbaited lines of live traps caught an equal number of Peromyscus. When one line was baited with rolled oats and peanut butter the efficiency of the traps was improved to the extent that the baited line captured more than twice as many mice as the unbaited line. It is concluded that for the species and habitat tested it is safe to make population calculations based on the assumption that the animals remain within their home ranges and do not tend to move into the trapped area because of the attraction of the trap bait.

  1. Environmental assessment requirements for live biological drugs.

    PubMed

    Sutton, Ann

    2008-02-01

    Marketing approval of biological products by the US Food and Drug Administration must comply with requirements of Code of Federal Regulations title 21 part 25, "Environmental Impact Considerations." An environmental impact statement is usually not required. Environmental assessment is required unless excluded. As naturally occurring substances, biological products qualify for categorical exclusion if manufacture and use do not significantly alter their concentration or distribution in the human environment. The manufacturing process and establishment descriptions in the license application should include enough detail to ensure that waste is controlled and inactivated. During clinical development of a live biotherapeutic product, data should be collected regarding the shedding of live organisms from treated patients. The ability of the live organism to persist in the environment should be assessed, and instructions for safe handling by health care providers and consumers should be incorporated into the package insert. PMID:18181713

  2. Recent synchronous radiation of a living fossil.

    PubMed

    Nagalingum, N S; Marshall, C R; Quental, T B; Rai, H S; Little, D P; Mathews, S

    2011-11-11

    Modern survivors of previously more diverse lineages are regarded as living fossils, particularly when characterized by morphological stasis. Cycads are often cited as a classic example, reaching their greatest diversity during the Jurassic-Cretaceous (199.6 to 65.5 million years ago) then dwindling to their present diversity of ~300 species as flowering plants rose to dominance. Using fossil-calibrated molecular phylogenies, we show that cycads underwent a near synchronous global rediversification beginning in the late Miocene, followed by a slowdown toward the Recent. Although the cycad lineage is ancient, our timetrees indicate that living cycad species are not much older than ~12 million years. These data reject the hypothesized role of dinosaurs in generating extant diversity and the designation of today's cycad species as living fossils. PMID:22021670

  3. Long lived NMR signal in bone

    NASA Astrophysics Data System (ADS)

    Zhang, Boyang; Lee, Jae-Seung; Khitrin, Anatoly; Jerschow, Alexej

    2013-06-01

    Solids and rigid tissues, such as bone, ligaments, and tendons, typically appear dark in MRI, which is due to the extremely short-lived proton nuclear magnetic resonance signals. This short lifetime is due to strong dipolar interactions between immobilized proton spins, which render it challenging to detect these signals with sufficient resolution and sensitivity. Here we show the possibility of exciting long-lived signals in cortical bone tissue with a signature consistent with that of bound water signals. It is further shown that dipolar coupling networks are an integral requirement for the excitation of these long-lived signals. The use of these signals could enhance the ability to visualize rigid tissues and solid samples with high resolution and sensitivity via MRI.

  4. Recent synchronous radiation of a living fossil.

    PubMed

    Nagalingum, N S; Marshall, C R; Quental, T B; Rai, H S; Little, D P; Mathews, S

    2011-11-11

    Modern survivors of previously more diverse lineages are regarded as living fossils, particularly when characterized by morphological stasis. Cycads are often cited as a classic example, reaching their greatest diversity during the Jurassic-Cretaceous (199.6 to 65.5 million years ago) then dwindling to their present diversity of ~300 species as flowering plants rose to dominance. Using fossil-calibrated molecular phylogenies, we show that cycads underwent a near synchronous global rediversification beginning in the late Miocene, followed by a slowdown toward the Recent. Although the cycad lineage is ancient, our timetrees indicate that living cycad species are not much older than ~12 million years. These data reject the hypothesized role of dinosaurs in generating extant diversity and the designation of today's cycad species as living fossils.

  5. Collecting live ant specimens (colony sampling).

    PubMed

    Smith, Chris R; Tschinkel, Walter R

    2009-07-01

    Because of the great diversity of ants, it is difficult to give a single protocol for the collection of live specimens. Ant body size can be very small or extremely large; the ants can be hard or soft, sting or spray toxic chemicals, live in the open or in hard-to-reach places; and colony size can range from tens of individuals to millions. Thus, collection techniques must be tailored to each particular species. In particular, caution must always be taken when dealing with stinging species, and symptoms and basic first-aid measures, especially for the treatment of anaphylactic shock, should be reviewed before beginning fieldwork. Nonetheless, many species are collectable as whole colonies. This protocol reviews some basic techniques for collecting ground-nesting species and describes how to collect whole live colonies (with queens), which are necessary for long-term laboratory studies and addressing questions of social organization and ecology. PMID:20147204

  6. Live cell imaging in Drosophila melanogaster.

    PubMed

    Parton, Richard M; Vallés, Ana Maria; Dobbie, Ian M; Davis, Ilan

    2010-04-01

    Although many of the techniques of live cell imaging in Drosophila melanogaster are also used by the greater community of cell biologists working on other model systems, studying living fly tissues presents unique difficulties with regard to keeping the cells alive, introducing fluorescent probes, and imaging through thick, hazy cytoplasm. This article outlines the major tissue types amenable to study by time-lapse cinematography and different methods for keeping the cells alive. It describes various imaging and associated techniques best suited to following changes in the distribution of fluorescently labeled molecules in real time in these tissues. Imaging, in general, is a rapidly developing discipline, and recent advances in imaging technology are able to greatly extend what can be achieved with live cell imaging of Drosophila tissues. As far as possible, this article includes the latest technical developments and discusses likely future developments in imaging methods that could have an impact on research using Drosophila. PMID:20360379

  7. Optical nanoscopy of a living cell

    NASA Astrophysics Data System (ADS)

    Ahluwalia, Balpreet S.; Wolfson, Deanna L.; Chuang, Frank Y. S.; Huser, Thomas

    2014-08-01

    Optical nanoscopy allows to study biological and functional processes of sub-cellular organelles. In structured illumination microscopy (SIM) the sample is illuminated with a grid-like interference pattern to encode higher spatial frequency information into observable Moiré patterns. By acquiring multiple images and a computation trick a superresolved image is obtained. SIM provides resolution enhancement of 2X in each axis as compared to conventional microscopes. For a visible light, SIM provides an optical resolution of 100 nm. The challenges associated with optical nanoscopy of a living cell are photo-toxicity, special dye requirements and artifacts due to cell movement. SIM works with conventional dyes and is a wide-field technique making it suitable for imaging living cells. In this work, we will discuss the opportunities and challenges of imaging living cells using SIM. Two applications of optical nanoscopy of living cells will be discussed; a) imaging of mitochondria in a keratinocyte cell and Optical microscopy based on fluorescence has emerged as a vital tool in modern bio-medical imaging and diagnosis. Super-resolution bio-imaging allows gathering information from sub-cellular organelles. In structured illumination microscopy (SIM) the sample is illuminated with a grid-like interference patterns to encode higher spatial frequencies information into observable images (Moiré fringes). A super-resolved image is then decoded using computational trick. In this work, we used SIM to acquired super-resolved optical images of mitochondria from a live keratinocyte cell (see Fig 1). SIM provides resolution enhancement of 2X in each axis and contrast enhancement of 8X on a projected image. Time-lapsed imaging was used to study the dynamics of mitochondria in a live cell.

  8. Producing a Live HDTV Program from Space

    NASA Technical Reports Server (NTRS)

    Grubbs, Rodney; Fontanot, Carlos; Hames, Kevin

    2007-01-01

    By the year 2000, NASA had flown HDTV camcorders on three Space Shuttle missions: STS-95, STS-93 and STS-99. All three flights of these camcorders were accomplished with cooperation from the Japanese space agency (then known as NASDA and now known as JAXA). The cameras were large broadcast-standard cameras provided by NASDA and flight certified by both NASA and NASDA. The high-definition video shot during these missions was spectacular. Waiting for the return of the tapes to Earth emphasized the next logical step: finding a way to downlink the HDTV live from space. Both the Space Shuttle and the International Space Station (ISS) programs were interested in live HDTV from space, but neither had the resources to fully fund the technology. Technically, downlinking from the ISS was the most effective approach. Only when the Japanese broadcaster NHK and the Japanese space agency expressed interest in covering a Japanese astronaut's journey to the ISS did the project become possible. Together, JAXA and NHK offered equipment, technology, and funding toward the project. In return, NHK asked for a live HDTV downlink during one of its broadcast programs. NASA and the ISS Program sought a US partner to broadcast a live HDTV program and approached the Discovery Channel. The Discovery Channel had proposed a live HDTV project in response to NASA's previous call for offers. The Discovery Channel agreed to provide addItional resources. With the final partner in place, the project was under way. Engineers in the Avionics Systems Division at NASA's Johnson Space Center (JSC) had already studied the various options for downlinking HDTV from the ISS. They concluded that the easiest way was to compress the HDTV so that the resulting data stream would "look" like a payload data stream. The flight system would consist of a professional HDTV camcorder with live HD-SDI output, an HDTV MPEG-2 encoder, and a packetizer/protocol converter.

  9. The Concept of Living and Non-Living Things in the World of Primary School Students in Turkey

    ERIC Educational Resources Information Center

    Topsakal, Unsal Umdu

    2010-01-01

    The purpose of this research is to reveal how the concepts of living and non-living things are in the world of the primary school (4th and 5th classes) students, what they remember when they are told about living and non-living things and what the characteristics of living and non-living things are according to them. The research is a descriptive…

  10. Arts Integration: An Exploration of the Dis/Connect between Policy and Live(d) Practice

    ERIC Educational Resources Information Center

    LaJevic, Lisa M.

    2009-01-01

    This dissertation explores the dis/connect between arts integration policy (i.e. written texts and curriculum documents) and the live(d) practice of teachers working with arts integration. Although previous studies have examined how arts integration is implemented in schools and how it affects student achievement, particularly standardized test…

  11. The Meaning of Work in the Lives of People Living with HIV Disease and AIDS.

    ERIC Educational Resources Information Center

    McReynolds, Connie J.

    2001-01-01

    Study focuses on the importance that work holds in the lives of people living with HIV disease and AIDS. Four categories were identified: providing access to affordable insurance and health care; providing a distraction from the disease; allowing one to be a contributing member of society; and serving as a measure of health. Implications for…

  12. Manipulability and Living/non-Living Category Effects on Object Identification

    ERIC Educational Resources Information Center

    Filliter, J.H.; McMullen, P.A.; Westwood, D.

    2005-01-01

    Object naming studies have generally observed that both normal and brain damaged individuals are faster and more accurate at identifying non-living objects than living objects (Humphreys, Riddoch, & Quinlan, 1988; Warrington & Shallice, 1984). However, a potential confounding variable, manipulability, has been present in past studies that may…

  13. [Ethical issues in live surgical demonstrations].

    PubMed

    Haruta, Seiichi

    2013-05-01

    Live surgical demonstrations are very useful educational tools for teaching skills, real-time problem solving, and how to manipulate new devices. However, such demonstrations may cause anxiety to surgeons because they are always aware of the commentator and attending physicians. Concentrating solely on the patient during an operative procedure is a medical ethical principle for surgeons. There are few advantages for patients in undergoing demonstration surgeries, except that they can expect the best surgeon in the field to operate. Therefore, surgeons should carefully control all aspects of live demonstrations, especially the indications, procedure selected, and results, to resolve ethical issues.

  14. Interactions between semiconductor nanowires and living cells.

    PubMed

    Prinz, Christelle N

    2015-06-17

    Semiconductor nanowires are increasingly used for biological applications and their small dimensions make them a promising tool for sensing and manipulating cells with minimal perturbation. In order to interface cells with nanowires in a controlled fashion, it is essential to understand the interactions between nanowires and living cells. The present paper reviews current progress in the understanding of these interactions, with knowledge gathered from studies where living cells were interfaced with vertical nanowire arrays. The effect of nanowires on cells is reported in terms of viability, cell-nanowire interface morphology, cell behavior, changes in gene expression as well as cellular stress markers. Unexplored issues and unanswered questions are discussed.

  15. Conservation: can we live better on less

    SciTech Connect

    Gore, R.

    1981-02-01

    Americans are looking for more-efficient ways to live and conduct their business without lowering their living standard. New building designs, intensive gardening, new energy sources, and vanpooling are among the pioneering efforts. Conservation also requires innovative ways to raise capital to handle nontraditional projects. A new industry of house doctors audits the energy efficiency of buildings and creates more-conserving designs and materials. Other industries will develop renewable and synthetic energy sources. Reports of changing attitudes and a growing interest in decentralized energy management are signs that conservation can become a way of life. (DCK)

  16. [Multifaceted body. 2. The lived body].

    PubMed

    Wykretowicz, H; Saraga, M; Bourquin, C; Stiefel, F

    2015-02-11

    The human body is the object upon which medicine is acting, but also lived reality, image, symbol, representation and the object of elaboration and theory. All these elements which constitute the body influence the way medicine is treating it. In this series of three articles, we address the human body from various perspectives: medical (1), phenomenological (2), psychosomatic and socio-anthropological (3). This second article distinguishes between the body as an object of knowledge or representation and the way the body is lived. This distinction which originates in phenomenological psychiatry aims to understand how the patient experiences his body and to surpass the classical somatic and psychiatric classifications. PMID:25895215

  17. Reassessing Medical Risk in Living Kidney Donors

    PubMed Central

    Kumar, Vineeta; Matas, Arthur J.

    2015-01-01

    The short- and long-term effects of unilateral nephrectomy on living donors have been important considerations for 60 years. Short-term risk is well established (0.03% mortality and <1% risk of major morbidity), but characterization of long-term risk is evolving. Relative to the general population, risk of mortality, ESRD, hypertension, proteinuria, and cardiovascular disease is comparable or lower. However, new studies comparing previous donors with equally healthy controls indicate increased risk of metabolic derangements (particularly involving calcium homeostasis), renal failure, and possibly, mortality. We discuss how these results should be interpreted and their influence on the practice of living donor kidney transplantation. PMID:25255922

  18. [Multifaceted body. 2. The lived body].

    PubMed

    Wykretowicz, H; Saraga, M; Bourquin, C; Stiefel, F

    2015-02-11

    The human body is the object upon which medicine is acting, but also lived reality, image, symbol, representation and the object of elaboration and theory. All these elements which constitute the body influence the way medicine is treating it. In this series of three articles, we address the human body from various perspectives: medical (1), phenomenological (2), psychosomatic and socio-anthropological (3). This second article distinguishes between the body as an object of knowledge or representation and the way the body is lived. This distinction which originates in phenomenological psychiatry aims to understand how the patient experiences his body and to surpass the classical somatic and psychiatric classifications.

  19. Informational biopolymer structure in early living forms.

    NASA Technical Reports Server (NTRS)

    Dayhoff, M. O.; Mclaughlin, P. J.; Barker, W. C.; Hunt, L. T.

    1972-01-01

    Some studies devoted to the search in various organisms for 'relics' of the biochemical nature of ancient organisms, preserved by the conservative nature of the evolutionary process in all living species, are reviewed. Investigations of five families of informational molecules constituting such 'relics' in very diverse organisms are reported. They include: cytochrome c, ferredoxin, trypsin, transfer ribonucleic acid (RNA), and 5S ribosomal RNA. It is shown that, even from these few informational molecules, some interesting inferences about early living organisms can be drawn.

  20. Imaging the Glycome in Living Systems

    PubMed Central

    Li, Boyangzi; Mock, Feiyan; Wu, Peng

    2016-01-01

    The glycome, the full complement of glycans that cells produce, is an attractive target for molecular imaging. Imaging of the glycome in living systems has recently been enabled via bioorthogonal chemical reporter-based approaches. In this chapter, we describe two approaches to introduce bioorthogonal chemical reporters (tags) onto cell surface fucosylated glycans and glycans bearing LacNAc disaccharides, respectively. The tagged glycans can then be conjugated to imaging probes via bioorthogonal click chemistry. Similar approaches can be extended to image other sectors of the glycome in living systems. PMID:22289465

  1. Colon-targeted delivery of live bacterial cell biotherapeutics including microencapsulated live bacterial cells

    PubMed Central

    Prakash, Satya; Malgorzata Urbanska, Aleksandra

    2008-01-01

    There has been an ample interest in delivery of therapeutic molecules using live cells. Oral delivery has been stipulated as best way to deliver live cells to humans for therapy. Colon, in particular, is a part of gastrointestinal (GI) tract that has been proposed to be an oral targeted site. The main objective of these oral therapy procedures is to deliver live cells not only to treat diseases like colorectal cancer, inflammatory bowel disease, and other GI tract diseases like intestinal obstruction and gastritis, but also to deliver therapeutic molecules for overall therapy in various diseases such as renal failure, coronary heart disease, hypertension, and others. This review provides a comprehensive summary of recent advancement in colon targeted live bacterial cell biotherapeutics. Current status of bacterial cell therapy, principles of artificial cells and its potentials in oral delivery of live bacterial cell biotherapeutics for clinical applications as well as biotherapeutic future perspectives are also discussed in our review. PMID:19707368

  2. Exploring the Living Planet with David Attenborough.

    ERIC Educational Resources Information Center

    Burgess, Jacquelin; Unwin, David

    1984-01-01

    In this interview David Attenborough, the celebrated natural history film maker and writer, talks about his highly successful television series, "The Living Planet." Devoted to the exposition of the world's ecosystems, the film represents a significant example of popular geographic education. (RM)

  3. Fermions Living in a Flat World

    SciTech Connect

    Jesus Anguiano-Galicia, Ma. de; Bashir, A.

    2006-09-25

    In a plane, parity transformation, which changes the sign of only one spatial coordinate, swaps the fermion fields living in two inequivalent representations. A parity invariant Lagrangian thus contains fields corresponding to both the representations. For such a Lagrangian, we show that we can also define a chiral symmetry.

  4. Living with Epilepsy--Not around It

    ERIC Educational Resources Information Center

    Apel, Laura

    2008-01-01

    This article presents an interview on Kevin Eggers, a 19-year-old college student from Seattle, Washington, who was diagnosed with epilepsy but had not let it prevent him from accomplishing his goals. As an Epilepsy Advocate, Kevin helps other teens and young adults realize that having a disability does not mean not living a normal and fulfilling…

  5. Live from Lincoln Center: MENC Teacher's Guide.

    ERIC Educational Resources Information Center

    Teaching Music, 1996

    1996-01-01

    Presents three lesson plans designed to work in conjunction with a live broadcast of the New York Philharmonic (may be available on videotape). The activities include identifying influences and components of works by Dvorak and Tchaikovsky. Includes a roster of the New York Philharmonic along with background material. (MJP)

  6. Community College Students Truly Live the Magic

    ERIC Educational Resources Information Center

    Shook, Stephanie

    2006-01-01

    This article talks about the Disney Theme Parks & Resorts College Program. The program attracts a variety of students each year from different backgrounds, major and career goals to the Walt Disney World Resort outside of Orlando, Florida, for a semester of living, learning and earning. The program has provided a foundation for thousands of…

  7. Synchrotron IR spectromicroscopy: chemistry of living cells.

    PubMed

    Holman, Hoi-Ying N; Bechtel, Hans A; Hao, Zhao; Martin, Michael C

    2010-11-01

    Advanced analytical capabilities of synchrotron IR spectromicroscopy meet the demands of modern biological research for studying molecular reactions in individual living cells. (To listen to a podcast about this article, please go to the Analytical Chemistry multimedia page at pubs.acs.org/page/ancham/audio/index.html.).

  8. Researching Graduates' Lived Experiences of Vocational Learning

    ERIC Educational Resources Information Center

    Leach, Tony

    2012-01-01

    The aim of this article is to exemplify the value of using a phenomenological approach when investigating graduates' lived experiences of vocational learning. For this study, qualitative data was obtained during a series of email interviews with 35 participants. As a group they are highly aspirational and, during their graduate studies, were…

  9. Bubbles in live-stranded dolphins.

    PubMed

    Dennison, S; Moore, M J; Fahlman, A; Moore, K; Sharp, S; Harry, C T; Hoppe, J; Niemeyer, M; Lentell, B; Wells, R S

    2012-04-01

    Bubbles in supersaturated tissues and blood occur in beaked whales stranded near sonar exercises, and post-mortem in dolphins bycaught at depth and then hauled to the surface. To evaluate live dolphins for bubbles, liver, kidneys, eyes and blubber-muscle interface of live-stranded and capture-release dolphins were scanned with B-mode ultrasound. Gas was identified in kidneys of 21 of 22 live-stranded dolphins and in the hepatic portal vasculature of 2 of 22. Nine then died or were euthanized and bubble presence corroborated by computer tomography and necropsy, 13 were released of which all but two did not re-strand. Bubbles were not detected in 20 live wild dolphins examined during health assessments in shallow water. Off-gassing of supersaturated blood and tissues was the most probable origin for the gas bubbles. In contrast to marine mammals repeatedly diving in the wild, stranded animals are unable to recompress by diving, and thus may retain bubbles. Since the majority of beached dolphins released did not re-strand it also suggests that minor bubble formation is tolerated and will not lead to clinically significant decompression sickness.

  10. Unwrapping Rap: A Literacy of Lived Experience.

    ERIC Educational Resources Information Center

    Brown, Stephen G.

    The adversarial forces of governmental censorship, freedom of expression, and capitalistic appropriation are engaged in an acrimonious debate over "Gangsta' Rap" that is being played out in the public spaces of popular culture. However, as a literacy of lived experience, Gangsta' Rap warrants critical investigation. Many postmodern theorists have…

  11. Successful Study Habits. Successful Living Skills.

    ERIC Educational Resources Information Center

    Oklahoma State Board of Vocational and Technical Education, Stillwater. Curriculum and Instructional Materials Center.

    This module on successful study habits is one of a series of modules designed to help teach students to become more self-sufficient in their personal and professional lives. This module contains teacher and student materials that are planned to allow students to identify areas that they need to improve in order to perform their best in school and…

  12. Developing a Research Agenda for Assisted Living

    ERIC Educational Resources Information Center

    Kane, Rosalie A.; Wilson, Keren Brown; Spector, William

    2007-01-01

    Purpose: We describe an approach to identifying knowledge gaps, research questions, and methodological issues for assisted living (AL) research. Design and Methods: We undertook an inventory of AL literature and research in progress and commissioned background papers critiquing knowledge on selected subtopics. With an advisory committee, we…

  13. Living Lightly: Energy Conservation in Housing.

    ERIC Educational Resources Information Center

    Bender, Tom

    This publication contains a series of papers which promote the concepts of energy conservation and offer safe and convenient ways of handling all aspects of our lives affected by energy without having to depend in any way on fossil fuels or nuclear power. These changes, which can be brought about in homes and in energy flows affected by the…

  14. Middle School Science Education for Sustainable Living

    ERIC Educational Resources Information Center

    Hudson, Peter

    2007-01-01

    This article discusses Science Education for Sustainable Living (SESL) and how SESL can empower middle school students. SESL implies educating people through scientific evidence toward justifiably potential solutions for sustaining life. In a school context, SESL may require students to understand current environmental issues, and this involves…

  15. Memory, Space and Time: Researching Children's Lives

    ERIC Educational Resources Information Center

    Moss, Dorothy

    2010-01-01

    This article discusses the research approach in "Pathways through Childhood", a small qualitative study drawing on memories of childhood. The research explores how wider social arrangements and social change influence children's everyday lives. The article discusses the way that the concepts of social memory, space and time have been drawn on to…

  16. Living in Japan. Intercultural Exchange Series. Revised.

    ERIC Educational Resources Information Center

    Hawkinson, Annie

    The guide provides a brief introduction to the culture and language of Japan, and is designed for visitors, students, and business travelers. It offers practical information on various aspects of daily living, including: money; banks; food; restaurants; hotels; tipping; postal and telecommunications services; transportation; shopping; health and…

  17. Foucault's Heterotopia and Children's Everyday Lives.

    ERIC Educational Resources Information Center

    McNamee, Sara

    2000-01-01

    Discusses Foucault's notion of "heterotopia"--real places but which exist unto themselves, such as a floating ship. Considers data on children's use of computer and video games to apply "heterotopia" to children's everyday social lives. Argues that childhood is subject to increasing boundaries, and that children create "other" spaces through…

  18. Definition and Classification of Assisted Living

    ERIC Educational Resources Information Center

    Zimmerman, Sheryl; Sloane, Philip D.

    2007-01-01

    Purpose: The purpose of this article is to discuss the benefits and limitations of, and considerations in, developing a typology of assisted living (AL). Design and Methods: We conducted a review and comparison of nine AL typologies drawn from the literature. Results: Typologies addressed matters related to the structure, process, population, and…

  19. Poor Americans: How the Poor White Live.

    ERIC Educational Resources Information Center

    Pilisuk, Marc; Pilisuk, Phyllis

    Contents of this book include the following essays which originally appeared in "Transaction" magazine: (1) "Poor Americans: an introduction," Marc Pilisuk and Phyllis Pilisuk; (2) "How the white poor live," Marc Pilisuk and Phyllis Pilisuk; (3) "The culture of poverty," Oscar Lewis; (4) "Life in Appalachia--the case of Hugh McCaslin," Robert…

  20. Children's Conceptions of Plants as Living Things.

    ERIC Educational Resources Information Center

    Wax, Naomi; Stavy, Ruth

    In this study, the attitudes of Israeli children aged 6 to 15 years were surveyed regarding their conceptions of plants as living things. It was desired to find out whether children consider plants to be alive, the knowledge differences between the different age groups in the study, how children classify plants according to biological criteria and…

  1. Going Prime Time with Live Chat Reference.

    ERIC Educational Resources Information Center

    Hoag, Tara J.; Cichanowicz, Edana McCaffrey

    2001-01-01

    Describes the development of the Suffolk Cooperative Library System's live, online chat reference service, a pilot project for public libraries in Suffolk County (New York). Topics include chat software selection; a virtual reference collection; marketing; funding; staffing; evaluation; expanded hours of service; email; and extracting data from…

  2. General Music and Children Living in Poverty

    ERIC Educational Resources Information Center

    McAnally, Elizabeth Ann

    2013-01-01

    A review of recent writing makes the case that children living in poverty (urban, rural, or other) are more in jeopardy now than ever. As teachers attest and research asserts, poverty affects children in profound, complicated, and lasting ways. However, the general music program is uniquely positioned to meet children’s needs, especially those…

  3. Allgemeinbildung: Readiness for Living in Risk Society

    ERIC Educational Resources Information Center

    Elmose, Steffen; Roth, Wolff-Michael

    2005-01-01

    Sociologists increasingly note that one lives in a risk society, characterized by the unpredictable consequences of techno-scientific innovation and production and by increasing complexity. Life in risk society, particularly in truly democratic societies, increasingly requires competencies not only to understand and change one's own circumstances…

  4. Live Scale Active Shooter Exercise: Lessons Learned

    ERIC Educational Resources Information Center

    Ervin, Randy

    2008-01-01

    On October 23, 2007, the Lake Land College Public Safety Department conducted a full-scale live exercise that simulated an active shooter and barricaded hostage. In this article, the author will emphasize what they learned, and how they intend to benefit from it. He will list the law enforcement issues and general issues they encountered, and then…

  5. Living the Work: Meditations on a Lark

    ERIC Educational Resources Information Center

    Neufeld, Jonathan A.

    2011-01-01

    It is widely assumed that there is a blanket norm requiring the performer to present the work "in the best light possible," and that the performer "make the ends of the work his own" or "live the work" in performance. Through careful consideration of a particular performance, I suggest that this is an inadequate conception of a performer's…

  6. Transitional Living Programs for Homeless Adolescents.

    ERIC Educational Resources Information Center

    Jarvis, Sara V.; Robertson, Robert M., Jr.

    This report presents a conceptual framework for developing, reviewing, and evaluating transitional living programs (TLPs) for homeless adolescents. It is designed to be used by those in the field who are or will be developing such programs. All TLPs share basic elements and each of these is described so that TLP providers can understand what their…

  7. Symposium: On Richard Shusterman's "Performing Live"

    ERIC Educational Resources Information Center

    Higgins, Kathleen Marie; Haskins, Casey; Shusterman, Richard

    2002-01-01

    In the introduction of "Performing Live", Richard Shusterman challenges the reader to decide of the book, "Do its diverse texts manifest an individual style, or at least a reasonably consistent one?" In reflecting on this question, Higgins answers, "Yes." For her, the essays in the book, although written over the course of a decade, do exhibit…

  8. Sustainable Living on the Tiwi Islands

    ERIC Educational Resources Information Center

    Burne, Cris; McKaige, Barbie

    2016-01-01

    This article reports on how the people of the Tiwi Islands (which lie in the Arafura Sea located off the coast of Darwin in Australia's Northern Territory) have carefully observed the rhythms and patterns of their country, developing a complex and precise way of living sustainably in their island environment. In 2015, the Tiwi people shared their…

  9. Food for Living Things, Teacher's Guide.

    ERIC Educational Resources Information Center

    Australian Council for Educational Research, Hawthorn.

    This teachers guide to "Food for Living Things," one of the units of the Junior Secondary Science Project prepared for use in Victorian (Australia) secondary schools, includes copies of all of the appropriate student materials in addition to information designed to assist the teacher develop the unit. This information includes hints on appropriate…

  10. Grandma, Mommy, and Me: Intergenerational Living

    ERIC Educational Resources Information Center

    Duffy, Roslyn

    2008-01-01

    Intergenerational living has become common. Single adults move back into the family home, due to finances or perhaps the need for child care. A 24-hour workforce, from nurses to grocery clerks to military deployment has turned "day"care needs into both "night and day" care crises. Unemployment, lack of health care or home foreclosures, as well as…

  11. Living Organisms for the Elementary Classroom.

    ERIC Educational Resources Information Center

    Hampton, Carolyn H.; Hampton, Carol D.

    This publication was prepared for elementary teachers and other local personnel responsible for providing, maintaining and using living organisms to enhance elementary science programs. The manual contains a foreword, general information, and an appendix. It gives information concerning equipment and supplies, establishing and maintaining an…

  12. Metro College for Living. Workshop Packet.

    ERIC Educational Resources Information Center

    Kreps, Alice Roelofs

    This packet contains information and materials for conducting a training workshop for working with adults who are developmentally disabled. The materials are specifically designed to train volunteer teachers in the College for Living (CFL) program, which supplements residential programs in and around Denver and aids institutions in orienting…

  13. Muslim Pupils' Lives Changed after Sept. 11

    ERIC Educational Resources Information Center

    Shah, Nirvi

    2011-01-01

    Since 9/11, the lives of some Muslim students, and those perceived to be Muslim, have changed across the country, shaped in part by the distrust and harassment Muslims have endured from fellow Americans. In the months immediately following the attacks, accounts of harassment of Muslim students mounted in the news media, as did efforts by Muslim…

  14. Southern Identity in "Southern Living" Magazine

    ERIC Educational Resources Information Center

    Lauder, Tracy

    2012-01-01

    A fantasy-theme analysis of the editors' letters in "Southern Living" magazine shows an editorial vision of valuing the past and showcasing unique regional qualities. In addition, a content analysis of the visual representation of race in the magazine's formative years and recent past validates that inhabitants of the region were portrayed…

  15. Living Green: Examining Sustainable Dorms and Identities

    ERIC Educational Resources Information Center

    Watson, Lesley; Johnson, Cathryn; Hegtvedt, Karen A.; Parris, Christie L.

    2015-01-01

    Purpose: The purpose of this study was to examine the effects of living in "green" dorms on students' environmentally responsible behaviors (ERBs), in concert with other factors, including individual identity and social context in the form of behavior modeling by peers. Design/methodology/approach: The sample of 243 consists of students…

  16. [Independent Living Skills: Guides and Curriculum Materials.

    ERIC Educational Resources Information Center

    Cosby, Jane

    The author offers curriculums, teaching guides, assessment instruments, and course descriptions for teaching independent living skills to handicapped adolescents and young adults. A guide on cooking and eating is designed to teach students how to use cooking utensils, cook healthy meals, plan nutritious meals, shop wisely, and budget food…

  17. Anterior Chamber Live Loa loa: Case Report.

    PubMed

    Kagmeni, G; Cheuteu, R; Bilong, Y; Wiedemann, P

    2016-01-01

    We reported a case of unusual intraocular Loa loa in a 27-year-old patient who presented with painful red eye. Biomicroscopy revealed a living and active adult worm in the anterior chamber of the right eye. After surgical extraction under local anesthesia, parasitological identification confirmed L. loa filariasis. PMID:27441005

  18. Fatherhood and Men's Lives at Middle Age

    ERIC Educational Resources Information Center

    Eggebeen, David J.; Dew, Jeffrey; Knoester, Chris

    2010-01-01

    This article uses data on 2,024 men who were followed through the third wave of the National Survey of Families and Households to examine the implications of fatherhood experiences for men's involvement in altruistic social activities at middle age. We find that middle-aged men (ages 45-65) who at some point in their lives become fathers are…

  19. An Overall County Quality of Living Index.

    ERIC Educational Resources Information Center

    Cleland, Charles L.; Lin, Y. N.

    A recent study of the quality of life in the rural South developed a weighted index of the overall quality of living based on subjective evaluations by rural residents of such concerns as the local government's quality, politics, political organizations, schools, educational system, churches, civic organizations, recreational facilities, job…

  20. Inquiry and Living History: Part III

    ERIC Educational Resources Information Center

    Coatney, Sharon; Smalley, Rachel

    2006-01-01

    The third article in this series featuring the Living History Program (see December 2005 and January 2006) looks at the life of Phillis Wheatley, the first published African American woman poet. Though little is known of her life as a slave in Boston, her published poetry is available today for people to read and enjoy. This study of Phillis…

  1. School Safety: Saving Lives with AEDs

    ERIC Educational Resources Information Center

    Slusser, Greg

    2012-01-01

    Automated external defibrillators (AEDs) on school and university campuses have saved many lives. Students, teachers and community members have been among the fortunate ones pulled from the brink of death. Medical studies validating the effectiveness of AEDs in schools and other public settings have been published in numerous medical journals.…

  2. Understanding the lives of older gay people.

    PubMed

    Kean, Reb

    2006-09-01

    Ten research articles were examined with the aim of increasing our understanding of the lives of older gay people. It is clear from the literature that nurses must not rely on stereotypes of older gay people to inform their professional practice. It is also important for nurses to appreciate that older homosexuals have different health and social care needs from their heterosexual contemporaries.

  3. The Musical Lives of Babies and Families.

    ERIC Educational Resources Information Center

    Fenichel, Emily, Ed.

    2002-01-01

    "Zero to Three" is a single focus bulletin of the National Center for Infants, Toddlers, and Families providing insight from multiple disciplines on the development of infants, toddlers, and their families. Noting that knowledge of childrens musical experiences in context is central to understanding childrens lives, and that reciprocally,…

  4. Classroom Diversity: Connecting Curriculum to Students' Lives.

    ERIC Educational Resources Information Center

    McIntyre, Ellen, Ed.; Rosebery, Ann, Ed.; Gonzalez, Norma, Ed.

    These papers examine the sociocultural approach to curriculum design, which provides minority and working class students with the same privileges that middle class students have (instruction that puts their knowledge and experiences at the heart of learning). It presents the theoretical framework for linking students' lives with curriculum and…

  5. Why do turtles live so long

    SciTech Connect

    Gibbons, J.W.

    1987-04-01

    Turtles appear to live longer than most other species of vertebrates, according to both maximal lifespans from zoo records and survivorship patterns in natural populations. Turtle longevity may reflect low metabolic activity, an absence of physiological and anatomical senility, a large investment in the adult's protective shell, and a life history with a long maturation period.

  6. Community Living in Three Wisconsin Counties.

    ERIC Educational Resources Information Center

    Taylor, Steven J.

    The site visit report describes community living programs for persons with severe disabilities in the Wisconsin counties of Dane, LaCrosse, and Columbia. The visit attempted to identify and document promising practices through interviews with administrators, officials and staff; observations of three foster homes, two small group homes, and two…

  7. Bubbles in live-stranded dolphins

    PubMed Central

    Dennison, S.; Moore, M. J.; Fahlman, A.; Moore, K.; Sharp, S.; Harry, C. T.; Hoppe, J.; Niemeyer, M.; Lentell, B.; Wells, R. S.

    2012-01-01

    Bubbles in supersaturated tissues and blood occur in beaked whales stranded near sonar exercises, and post-mortem in dolphins bycaught at depth and then hauled to the surface. To evaluate live dolphins for bubbles, liver, kidneys, eyes and blubber–muscle interface of live-stranded and capture-release dolphins were scanned with B-mode ultrasound. Gas was identified in kidneys of 21 of 22 live-stranded dolphins and in the hepatic portal vasculature of 2 of 22. Nine then died or were euthanized and bubble presence corroborated by computer tomography and necropsy, 13 were released of which all but two did not re-strand. Bubbles were not detected in 20 live wild dolphins examined during health assessments in shallow water. Off-gassing of supersaturated blood and tissues was the most probable origin for the gas bubbles. In contrast to marine mammals repeatedly diving in the wild, stranded animals are unable to recompress by diving, and thus may retain bubbles. Since the majority of beached dolphins released did not re-strand it also suggests that minor bubble formation is tolerated and will not lead to clinically significant decompression sickness. PMID:21993505

  8. Adult and Family Living. Teacher Edition.

    ERIC Educational Resources Information Center

    Oklahoma State Dept. of Vocational and Technical Education, Stillwater. Curriculum and Instructional Materials Center.

    This curriculum guide for teachers is designed for use with 11th- and 12th-grade students who have had no more than 1 year of vocational home economics. It focuses on providing young adults with the knowledge and skills they need for healthy and positive adult and family lives. It includes 27 units in 8 sections as follows: (1) personal…

  9. Live from Westminster: Broadcasting the British Parliament.

    ERIC Educational Resources Information Center

    Scott, Peter Hardiman

    1978-01-01

    Describes live radio broadcasting from the chambers of the British Parliament. After 55 years of campaigning by broadcasters and Members of Parliament, and following experimental broadcasting by the BBC and Independent Local Radio, service was installed in April 1978. Initial experimentation, current procedures, and implications for television…

  10. Developing a Living Archive of Aboriginal Languages

    ERIC Educational Resources Information Center

    Bow, Catherine; Christie, Michael; Devlin, Brian

    2014-01-01

    The fluctuating fortunes of Northern Territory bilingual education programs in Australian languages and English have put at risk thousands of books developed for these programs in remote schools. In an effort to preserve such a rich cultural and linguistic heritage, the Living Archive of Aboriginal Languages project is establishing an open access,…

  11. Living Arrangements: A Closer Look at Families.

    ERIC Educational Resources Information Center

    University of South Florida, Tampa. Louis de la Parte Florida Mental Health Inst.

    This Kids Count pamphlet provides information on the living arrangements of Florida's children, focusing on family types. Drawing on information from the 1997 National Survey of America's Families, the Current Population Survey, and the National Center for Health Statistics, the pamphlet presents information on changes in America's families,…

  12. Live @ The Exploratorium: Origins. Final Evaluation Report

    ERIC Educational Resources Information Center

    Spielvogel, Robert; Chang, Han-Hua

    2004-01-01

    In the summer of 2000, Education Development Center's Center for Children and Technology (CCT) was contracted to evaluate the LIVE @ THE EXPLORATORIUM: Origins project as part of the Exploratorium's three-year funding from the National Science Foundation. Evaluation planning and work began in August of 2000 and continued through December 2003.…

  13. Improving Health Care for Assisted Living Residents

    ERIC Educational Resources Information Center

    Kane, Robert L.; Mach, John R., Jr.

    2007-01-01

    Purpose: The purpose of this article is to explore how medical care is delivered to older people in assisted living (AL) settings and to suggest ways for improving it. Design and Methods: We present a review of the limited research available on health care for older AL residents and on building testable models of better ways to organize primary…

  14. Adaptive Dialogue Systems for Assistive Living Environments

    ERIC Educational Resources Information Center

    Papangelis, Alexandros

    2013-01-01

    Adaptive Dialogue Systems (ADS) are intelligent systems, able to interact with users via multiple modalities, such as speech, gestures, facial expressions and others. Such systems are able to make conversation with their users, usually on a specific, narrow topic. Assistive Living Environments are environments where the users are by definition not…

  15. Living Sculptures: Performance Art in the Classroom

    ERIC Educational Resources Information Center

    Pembleton, Matthew; LaJevic, Lisa

    2014-01-01

    What does an introduction to and engagement in performance art offer K-12 students? In this article, we respond to this question by proposing a lesson inspired by the artmaking practices of the contemporary artist Erwin Wurm. Performance art can be defined as any form of work that combines the artist's body and a live-action event with or…

  16. Facts and Fancies On High Rise Living

    PubMed Central

    Wilson, Murray; Steinbach, Andrew

    1973-01-01

    The authors, after reviewing various findings and opinions about the effects of high rise living on each segment of the population, state their own findings from a six-year practice in a downtown Toronto complex of single family units and apartments. They show that the pathology of patients does not differ markedly in either group. PMID:20468903

  17. Anterior Chamber Live Loa loa: Case Report

    PubMed Central

    Kagmeni, G.; Cheuteu, R.; Bilong, Y.; Wiedemann, P.

    2016-01-01

    We reported a case of unusual intraocular Loa loa in a 27-year-old patient who presented with painful red eye. Biomicroscopy revealed a living and active adult worm in the anterior chamber of the right eye. After surgical extraction under local anesthesia, parasitological identification confirmed L. loa filariasis. PMID:27441005

  18. Living kidney donation: outcomes, ethics, and uncertainty.

    PubMed

    Reese, Peter P; Boudville, Neil; Garg, Amit X

    2015-05-16

    Since the first living-donor kidney transplantation in 1954, more than half a million living kidney donations have occurred and research has advanced knowledge about long-term donor outcomes. Donors in developed countries have a similar life expectancy and quality of life as healthy non-donors. Living kidney donation is associated with an increased risk of end-stage renal disease, although this outcome is uncommon (<0·5% increase in incidence at 15 years). Kidney donation seems to elevate the risks of gestational hypertension and pre-eclampsia. Many donors incur financial expenses due to factors such as lost wages, need for sick days, and travel expenses. Yet, most donors have no regrets about donation. Living kidney donation is practised ethically when informed consent incorporates information about risks, uncertainty about outcomes is acknowledged when it exists, and a donor's risks are proportional to benefits for the donor and recipient. Future research should determine whether outcomes are similar for donors from developing countries and donors with pre-existing conditions such as obesity. PMID:26090646

  19. Living kidney donation: outcomes, ethics, and uncertainty.

    PubMed

    Reese, Peter P; Boudville, Neil; Garg, Amit X

    2015-05-16

    Since the first living-donor kidney transplantation in 1954, more than half a million living kidney donations have occurred and research has advanced knowledge about long-term donor outcomes. Donors in developed countries have a similar life expectancy and quality of life as healthy non-donors. Living kidney donation is associated with an increased risk of end-stage renal disease, although this outcome is uncommon (<0·5% increase in incidence at 15 years). Kidney donation seems to elevate the risks of gestational hypertension and pre-eclampsia. Many donors incur financial expenses due to factors such as lost wages, need for sick days, and travel expenses. Yet, most donors have no regrets about donation. Living kidney donation is practised ethically when informed consent incorporates information about risks, uncertainty about outcomes is acknowledged when it exists, and a donor's risks are proportional to benefits for the donor and recipient. Future research should determine whether outcomes are similar for donors from developing countries and donors with pre-existing conditions such as obesity.

  20. Building Resilient Lives: New Zealand Health Camps

    ERIC Educational Resources Information Center

    Rose, Kirsten

    2009-01-01

    Sometimes it takes a child to raise a whole village. Young people have the gifts and talents to raise their villages. They lack only the confidence of their villages in them to do it. At Te Puna Whaiora Children's Health Camps in New Zealand, the approach to helping families reach their potential is radically changing the lives of many children…