In vitro and in vivo development of mice morulae after storage in non-frozen conditions

BACKGROUND: Interchange of genetically modified (GM) mice between laboratories using embryos provides several advantages. Not only is transport stress avoided, but also the health status of the recipient colony is not compromised. Embryos do not need to be shipped in frozen stage, which requires expensive packaging in addition to a certain degree of expertise in order to freeze and thaw them correctly. The aim of this study was to examine different storage conditions and their effect on embryo viability in order to establish the feasibility of practical, non-frozen conditions for embryo shipment. METHODS: Mouse morulae developed in vivo (collected from donors 2.5d post coitum) or in vitro (zygotes cultured until morulae stage) were stored, combining two different media (KSOMeq or KSOM-H) and temperatures (4 degrees C, 15 degrees C and 37 degrees C) throughout 24 or 48 hours. After storage in vitro viability was assessed determining percentage of development to blastocyst and total cell number. In vivo viability was determined based on the number of implantations and living fetuses after embryo transfer of stored embryos. The storage effect at the molecular level was assessed by studying a gene pool involved in early development by quantitative RT-PCR. RESULTS: In vivo-produced morulae stored for 24 hours did not show differences in development up to the blastocyst stage, regardless of the storage type. Even though a decrease in the total cell number in vivo was observed, embryo development after embryo transfer was not affected. All 24 hour storage conditions tested provided a similar number of implantations and fetuses at day 14 of pregnancy. Morulae obtained from in vitro embryo culture collected at the 1-cell stage showed a decreased ability to develop to blastocyst after 24 hours of storage at 15degrees C both in KSOMeq and KSOM-H. Concomitantly, a significant decrease of embryo implantation rates after transfer to recipients was also found. In order to further characterize the effect of non-frozen storage combining a molecular approach with the ordinary in vitro culture evaluation, embryos collected at the morula stage were submitted to the same storage conditions described throughout 48 hours. In vitro culture of those embryos showed a significant decrease in their developmental rate to blastocyst in both KSOMeq and KSOM-H at 15degrees C, which also affected the total number of cells. Gene transcription studies confirmed significant alterations in retrotransposons (Erv4 and Iap) after 48 h of storage at 15degrees C. CONCLUSIONS: Our results show that both KSOMeq and KSOM-H can be equally used, and that several temperature conditions allow good survival rates in vitro and in vivo. Some of these storage conditions can substitute freezing in order to maintain embryo viability for 24–48 hours, providing a reliable and less demanding technical alternative for embryo interchanges

Aproximación a la producción y liderazgo de las mujeres en la geografía española

The article presents the panorama of women representation in Spanish geography through the analysis of different indicators of scientific production (articles, books, and book chapters) and leadership (head of research groups, presence on scientific committees of journals and coordination of books). The study is fundamentally based on the exploitation of information from the Dialnet bibliographic portal. The analysis of leadership has been complemented with other sources of information related to research group coordination and participation in editorial committees of scientific journals. The results show a greater number of male geographers than female geographers belonging to the three geographic areas (Physical, Regional and Human) though there are slightly differences regarding the institution and the area of knowledge. Results also highlight the greater production of the males and an imbalance panorama regarding leadership.El artículo muestra el panorama de la representación de las mujeres en la geografía española a través de distintos indicadores relacionados con la producción científica (artículos, libros y capítulos de libros) y el liderazgo (responsables de grupos de investigación, presencia en los comités científicos de revistas y coordinación de libros). El estudio se basa fundamentalmente en la explotación de la información del portal bibliográfico Dialnet, cuyo repositorio ha servido de aproximación para el análisis de la producción científica en la geografía española de hombres y mujeres. El análisis del liderazgo ha sido complementado con otras fuentes disponibles en internet sobre dirección de grupos de investigación y participación en comités editoriales de revistas científicas. Los resultados muestran un mayor número de autores que de autoras adscritos a las tres áreas de conocimiento de la Geografía (Física, Regional y Humana), aunque con matices por institución y áreas, una mayor trayectoria y profusa producción de aquellos, así como un panorama desigual en lo relativo al desempeño actual en puestos de liderazgo.L'article présente le panorama des femmes dans la géographie espagnole à travers la production scientifique (articles, livres et chapitres de livres) et le leadership (responsables de groupes de recherche, présence dans les comités scientifiques de revues et coordination d'ouvrages). L'étude repose fondamentalement sur l'exploitation des informations du portail bibliographique Dialnet, qui ont été traitées via le programme d'analyse de texte Atlas.ti. L'analyse du leadership a été complétée par d'autres sources disponibles sur Internet sur la gestion des groupes de recherche et la participation aux comités éditoriaux des revues. Les résultats montrent un plus grand nombre de géographes hommes que de femmes géographes, une plus grande production des premiers et un plus grand leadership

The proximal promoter region of mTert is sufficient to regulate telomerase activity in ES cells and transgenic animals

BACKGROUND: The reverse transcriptase of telomerase (Tert) controls telomerase activity maintaining the end of linear chromosomes in eukaryotic cells. Telomerase function is highly active in undifferentiated multipotent stem cells, decreases with cell differentiation and is generally absent from most somatic cells in the adult. Its absence is responsible of telomeres shortening in such somatic cells. Using an in vivo transgenic model and an in vitro culture differentiation of adult stem cells, we examined the elements of the mouse Tert (mTert) promoter that control telomerase activity. RESULTS: Three constructs comprising 1, 2 or 5 kb of the mTert promoter sequence coupled to the coding sequence of the green fluorescent protein (EGFP) were electroporated into embryonic stem (ES) cells. Transformed ES cells were able to mimic the expected mTert expression, which was associated to green fluorescence. One and 5 kb promoter produced the higher expression of EGFP, on ES cells. When ES cells were allowed to differentiate to embryoid bodies and to other cell types, they lost gradually the expression of mTert-EGFP as consequence of differentiation. No differences were found among the three constructs analyzed. We then generated transgenic mice with the three constructs. Expression of the reporter gene was monitored by reverse transcription-PCR analysis and EGFP visualization. The mRNA expression of the three constructs was lower than the endogenous mTert, but mimicked the endogenous mTert transcription pattern; however, no fluorescent expression of EGFP was detected in adult tissues. EGFP expression of the three constructs was visualized at the blastocysts stage and in new ES cells generated from them; in the germinal ring of E13 dpc foetuses; in ES-like colonies and in germinal stem cells generated from neonatal and adult testis cells; and in neuroesferes generated from E14 dpc foetuses' brain cells. CONCLUSION: The 1 kb promoter upstream of the initiating ATG codon of mTert contains all the regulatory elements to control telomerase expression in ES cells during in vitro loss of pluripotency. The transgenic mouse lines generated represent an appropriate system to analyze the expression of mouse Tert gene under physiological condition and during establishment of stem cell lines generated from embryonic or adult tissues

Racism: A Teenagers\u27 Perspective Results of Preliminary Research from Madrid, Spain

In mid-June, 2005, the members of the INTER Center received a collaboration proposal from FETE-UGT8, with the objective of carrying out a brief exploratory study on the perceptions and experiences that young people and adolescents, mainly immigrants, have concerning possible experiences of discrimination and racism in their immediate surroundings. The initial objectives of the project were expanded due to the dynamics of the project itself. New focuses of attention and social, educational and personal dynamics, which can condition to a certain extent the experiences that immigrant adolescents undergo, were detected. The project initially consisted of a series of interviews with adolescents between 12 and 18 years of age. A total of 20 interviews were carried out, some of them group interviews. We consider this project focus, although positive, to be limited, and we think that it would have been necessary to broaden the sample and consider other people in the young people’s social and family environment (family members, friends, classmates, professors, all of them of different origins, including the host country)

AVALIAÇÃO DA SUSTENTABILIDADE NA GESTÃO DE CLÍNICAS DENTÁRIAS. REVISÃO DA LITERATURA

Sustainability in dentistry is based on the need to integrate responsible environmental practices within the dental field, and processes that minimize negative environmental impact, promoting public health and economic efficiency. This integration ranges from the reduction of carbon emissions to proper waste management and optimization of natural resources. Objectives: The objective of this narrative review is to evaluate current strategies and practices in sustainable dentistry to identify effective methods that help reduce the environmental footprint of dental clinics. Methods: The research was structured based on a search tree PubMed, Google Scholar, ProQuest, Scopus. The languages: English, Portuguese and Spanish were used as limiters; free access articles, any type of research study. We established a secondary selection based on the title and abstract according to the objective of the study. Results: The initial search showed 6,092 studies whose title was related to the search patterns, 6,025 articles were excluded based on their title and abstract, resulting in 67 relevant articles. After reading, 24 articles that did not meet the inclusion criteria were excluded: Thus, 42 articles were finally selected for the present review. Conclusions:  The implementation of sustainable practices in dental clinics not only seeks to reduce the environmental footprint and promote operational efficiency, but also strengthens the corporate social responsibility of the sector. These actions are essential to moving towards a healthy and sustainable healthcare environment, aligned with global environmental conservation principles.La sostenibilidad en odontología se fundamenta en la necesidad de integrar prácticas ambientales responsables dentro del campo dental, y procesos que minimicen el impacto ambiental negativo, promoviendo la salud pública y la eficiencia económica. Esta integración abarca desde la reducción de emisiones de carbono hasta la gestión adecuada de residuos y la optimización del uso de recursos naturales. Objetivos: El objetivo de esta revisión narrativa es evaluar las estrategias y prácticas actuales en odontología sustentable para identificar métodos efectivos que ayuden a reducir la huella ambiental de las clínicas dentales. Métodos: La investigación fue estructurada a partir de un árbol de búsqueda (PubMed, Google Académico, ProQuest, Scopus). Se utilizaron como limitadores los idiomas: Inglés, Portugués y Español; artículos de libre acceso, cualquier tipología de estudio de investigación. Establecimos una selección secundaria a partir del título y resumen de los artículos de acuerdo al objetivo del estudio. Resultados: La búsqueda inicial mostró 6.092 estudios cuyo título estaba relacionado con los patrones de búsqueda, se excluyeron 6.025 artículos en función de su título y resumen, resultando 67 artículos potencialmente relevantes. Luego de la lectura, fueron excluidos 24 artículos que no cumplían con los criterios de inclusión: Así, finalmente se seleccionaron 42 artículos para la presente revisión. La sostenibilidad en odontología se fundamenta en la necesidad de integrar prácticas ambientales responsables dentro del campo dental, y procesos que minimicen el impacto ambiental negativo, promoviendo la salud pública y la eficiencia económica. Esta integración abarca desde la reducción de emisiones de carbono hasta la gestión adecuada de residuos y la optimización del uso de recursos naturales. Objetivos: El objetivo de esta revisión narrativa es evaluar las estrategias y prácticas actuales en odontología sustentable para identificar métodos efectivos que ayuden a reducir la huella ambiental de las clínicas dentales. Métodos: La investigación fue estructurada a partir de un árbol de búsqueda (PubMed, Google Académico, ProQuest, Scopus). Se utilizaron como limitadores los idiomas: Inglés, Portugués y Español; artículos de libre acceso, cualquier tipología de estudio de investigación. Establecimos una selección secundaria a partir del título y resumen de los artículos de acuerdo al objetivo del estudio. Resultados: La búsqueda inicial mostró 6.092 estudios cuyo título estaba relacionado con los patrones de búsqueda, se excluyeron 6.025 artículos en función de su título y resumen, resultando 67 artículos potencialmente relevantes. Luego de la lectura, fueron excluidos 24 artículos que no cumplían con los criterios de inclusión: Así, finalmente se seleccionaron 42 artículos para la presente revisión. A sustentabilidade na medicina dentária baseia-se na necessidade de integrar práticas ambientais responsáveis ​​na área dentária e processos que minimizem o impacto ambiental negativo, promovendo a saúde pública e a eficiência económica. Esta integração vai desde a redução das emissões de carbono até a gestão adequada dos resíduos e a otimização da utilização dos recursos naturais. Objetivos: O objetivo desta revisão narrativa é avaliar estratégias e práticas atuais em odontologia sustentável para identificar métodos eficazes que ajudem a reduzir a carga ambiental das práticas odontológicas. Métodos: A busca foi estruturada com base em uma árvore de busca (PubMed, Google Scholar, ProQuest, Scopus). Serão utilizados como idiomas limitantes os seguintes idiomas: inglês, português e espanhol; Artigos de acesso gratuito, qualquer tipo de pesquisa. Estabelecemos uma seleção secundária baseada no título e resumo dos artigos de acordo com o objetivo do estudo. Resultados: A busca inicial apresentou 6.092 estudos cujo título estava relacionado aos patrocinadores da busca, excluindo 6.025 artigos por título e resumo, resultando em 67 artigos potencialmente relevantes. Durante a leitura foram excluídos 24 artigos que não atendiam aos critérios de inclusão: assim, foram finalmente selecionados 42 artigos para a presente análise. Conclusões: A implementação de práticas sustentáveis ​​nas clínicas odontológicas presenciais busca reduzir a poluição ambiental e promover a eficiência operacional, o que também fortalece a responsabilidade social corporativa do setor. Estas ações são essenciais para promover um ambiente de saúde mais saudável e sustentável, alinhado aos princípios globais de conservação ambiental

Precursors of fatty alcohols in the ISM: Discovery of n-propanol

Theories on the origins of life propose that early cell membranes were synthesized from amphiphilic molecules simpler than phospholipids such as fatty alcohols. The discovery in the interstellar medium (ISM) of ethanolamine, the simplest phospholipid head group, raises the question whether simple amphiphilic molecules are also synthesized in space. We investigate whether precursors of fatty alcohols are present in the ISM. For this, we have carried out a spectral survey at 7, 3, 2 and 1 mm toward the Giant Molecular Cloud G+0.693-0.027 located in the Galactic Center using the IRAM 30m and Yebes 40m telescopes. Here, we report the detection in the ISM of the primary alcohol n-propanol (in both conformers Ga-n-C3H7OH and Aa-n-C3H7OH), a precursor of fatty alcohols. The derived column densities of n-propanol are (5.5+-0.4)x10^13 cm^-2 for the Ga conformer and (3.4+-0.3)x10^13 cm^-2 for the Aa conformer, which imply molecular abundances of (4.1+-0.3)x10^-10 for Ga-n-C3H7OH and of (2.5+-0.2)x10^-10 for Aa-n-C3H7OH. We also searched for the AGa conformer of n-butanol (AGa-n-C4H9OH) without success yielding an upper limit to its abundance of <4.1x10^-11. The inferred CH3OH:C2H5OH:C3H7OH:C4H9OH abundance ratios go as 1:0.04:0.006:<0.0004 toward G+0.693-0.027, i.e. they decrease roughly by one order of magnitude for increasing complexity. We also report the detection of both syn and anti conformers of vinyl alcohol, with column densities of (1.11+-0.08)x10^14 cm^-2 and (1.3+-0.4)x10^13 cm^-2, and abundances of (8.2+-0.6)x10^-10 and (9.6+-3.0)x10^-11, respectively. The detection of n-propanol, together with the recent discovery of ethanolamine in the ISM, opens the possibility that precursors of lipids according to theories of the origin of life, could have been brought to Earth from outer space.Comment: 15 pages, 10 figures, accepted for A&

Sheep and Goat BSE Propagate More Efficiently than Cattle BSE in Human PrP Transgenic Mice

A new variant of Creutzfeldt Jacob Disease (vCJD) was identified in humans and linked to the consumption of Bovine Spongiform Encephalopathy (BSE)-infected meat products. Recycling of ruminant tissue in meat and bone meal (MBM) has been proposed as origin of the BSE epidemic. During this epidemic, sheep and goats have been exposed to BSE-contaminated MBM. It is well known that sheep can be experimentally infected with BSE and two field BSE-like cases have been reported in goats. In this work we evaluated the human susceptibility to small ruminants-passaged BSE prions by inoculating two different transgenic mouse lines expressing the methionine (Met) allele of human PrP at codon 129 (tg650 and tg340) with several sheep and goat BSE isolates and compared their transmission characteristics with those of cattle BSE. While the molecular and neuropathological transmission features were undistinguishable and similar to those obtained after transmission of vCJD in both transgenic mouse lines, sheep and goat BSE isolates showed higher transmission efficiency on serial passaging compared to cattle BSE. We found that this higher transmission efficiency was strongly influenced by the ovine PrP sequence, rather than by other host species-specific factors. Although extrapolation of results from prion transmission studies by using transgenic mice has to be done very carefully, especially when human susceptibility to prions is analyzed, our results clearly indicate that Met129 homozygous individuals might be susceptible to a sheep or goat BSE agent at a higher degree than to cattle BSE, and that these agents might transmit with molecular and neuropathological properties indistinguishable from those of vCJD. Our results suggest that the possibility of a small ruminant BSE prion as vCJD causal agent could not be ruled out, and that the risk for humans of a potential goat and/or sheep BSE agent should not be underestimated

Effectiveness of Fosfomycin for the Treatment of Multidrug-Resistant Escherichia coli Bacteremic Urinary Tract Infections

IMPORTANCE The consumption of broad-spectrum drugs has increased as a consequence of the spread of multidrug-resistant (MDR) Escherichia coli. Finding alternatives for these infections is critical, for which some neglected drugs may be an option. OBJECTIVE To determine whether fosfomycin is noninferior to ceftriaxone or meropenem in the targeted treatment of bacteremic urinary tract infections (bUTIs) due to MDR E coli. DESIGN, SETTING, AND PARTICIPANTS This multicenter, randomized, pragmatic, open clinical trial was conducted at 22 Spanish hospitals from June 2014 to December 2018. Eligible participants were adult patients with bacteremic urinary tract infections due to MDR E coli; 161 of 1578 screened patients were randomized and followed up for 60 days. Data were analyzed in May 2021. INTERVENTIONS Patients were randomized 1 to 1 to receive intravenous fosfomycin disodium at 4 g every 6 hours (70 participants) or a comparator (ceftriaxone or meropenem if resistant; 73 participants) with the option to switch to oral fosfomycin trometamol for the fosfomycin group or an active oral drug or pa renteral ertapenem for the comparator group after 4 days. MAIN OUTCOMES AND MEASURES The primary outcome was clinical and microbiological cure (CMC) 5 to 7 days after finalization of treatment; a noninferiority margin of 7% was considered. RESULTS Among 143 patients in the modified intention-to-treat population (median [IQR] age, 72 [62-81] years; 73 [51.0%] women), 48 of 70 patients (68.6%) treated with fosfomycin and 57 of 73 patients (78.1%) treated with comparators reached CMC (risk difference, -9.4 percentage points; 1-sided 95% CI, -21.5 to infinity percentage points; P = .10). While clinical or microbiological failure occurred among 10 patients (14.3%) treated with fosfomycin and 14 patients (19.7%) treated with comparators (risk difference, -5.4 percentage points; 1-sided 95% CI. -infinity to 4.9; percentage points; P = .19), an increased rate of adverse event-related discontinuations occurred with fosfomycin vs comparators (6 discontinuations [8.5%] vs 0 discontinuations; P = .006). In an exploratory analysis among a subset of 38 patients who underwent rectal colonization studies, patients treated with fosfomycin acquired a new ceftriaxone-resistant or meropenem-resistant gram-negative bacteria at a decreased rate compared with patients treated with comparators (0 of 21 patients vs 4 of 17 patients [23.5%]; 1-sided P = .01). CONCLUSIONS AND RELEVANCE This study found that fosfomycin did not demonstrate noninferiority to comparators as targeted treatment of bUTI from MDR E coli; this was due to an increased rate of adverse event-related discontinuations. This finding suggests that fosfomycin may be considered for selected patients with these infections

Prevalencia, características clínicas y evolución de la anemia hemolítica inmune en el Hospital Nacional Dos de mayo desde enero 2000 hasta enero 2005

Establece la prevalencia, características clínicas y evolución de las anemias hemolíticas inmunes en el Hospital Nacional Dos de Mayo de Lima, Perú durante el periodo enero 2000 - enero 2005. Se realizó un estudio descriptivo observacional, retrospectivo y transversal con el total de pacientes con diagnóstico de anemia hemolítica inmune mayores de 14 años que presentaban evidencia de hemólisis y prueba de Coombs directa positiva, registrados en los archivos del Servicio de Hematología de la institución. El cuadro clínico se caracterizó porque casi todos lo pacientes presentaron palidez, acompañada de uno o más signos o síntomas, siendo el más frecuente la palidez e ictericia (23,5%). El nivel de anemia más frecuente fue la severa (55,9%), seguida por la moderada (41,2%) y la leve (2,9%). Predominó la anemia hemolítica autoinmune causada por anticuerpos calientes (85,3%) y la IgG fue el principal patrón antiglobulínico (50%), seguido por la IgG C3d (20,6%) y la IgM (11,8%). La principal enfermedad secundaria fue el lupus eritematoso sistémico (53,8%), aunque también se observó dos casos de cirrosis biliar primaria (5,9%) y dos de síndrome de Evans (5,9%). El 26,5% de pacientes eran refractarios al tratamiento, el 64,7% (n = 22) respondió a los corticoides y el 8,8% presentó cura espontánea. La remisión clínica, serológica y hematológica con la corticoterapia fue más frecuente antes los 6 meses de tratamiento. Se concluye que la anemia hemolítica autoinmune es una enfermedad rara causada principalmente por anticuerpos calientes que responde normalmente a la terapia convencional con corticoesteroides, aunque su diagnóstico debería alertar sobre la posibilidad de una enfermedad sistémica asociada.Trabajo académic