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1.
PLoS Genet ; 18(2): e1010060, 2022 02.
Artigo em Inglês | MEDLINE | ID: mdl-35180218

RESUMO

The increasing prevalence of insecticide resistance and the ongoing global burden of vector-borne diseases have encouraged new efforts in mosquito control. For Aedes aegypti, the most important arboviral vector, integration rates achieved in Cas9-based knock-ins so far have been rather low, highlighting the need to understand gene conversion patterns and other factors that influence homology-directed repair (HDR) events in this species. In this study, we report the effects of sequence mismatches or donor template forms on integration rates. We found that modest sequence differences between construct homology arms [DNA sequence in the donor template which resembles the region flanking the target cut] and genomic target comprising 1.2% nucleotide dissimilarity (heterology) significantly reduced integration rates. While most integrations (59-88%) from plasmid templates were the result of canonical [on target, perfect repair] HDR events, no canonical events were identified from other donor types (i.e. ssDNA, biotinylated ds/ssDNA). Sequencing of the transgene flanking region in 69 individuals with canonical integrations revealed 60% of conversion tracts to be unidirectional and extend up to 220 bp proximal to the break, though in three individuals bidirectional conversion of up to 725 bp was observed.


Assuntos
Sistemas CRISPR-Cas , Culicidae , Animais , Culicidae/genética , Reparo do DNA/genética , Genoma , Humanos , Mosquitos Vetores/genética
2.
Insect Mol Biol ; 2024 Aug 11.
Artigo em Inglês | MEDLINE | ID: mdl-39129057

RESUMO

Anopheles stephensi Liston, 1901 (Diptera: culicidae) is a competent vector of Plasmodium falciparum (Haemosporida: plasmodiidae) malaria, and its expansion in the African continent is of concern due to its viability in urban settings and resistance to insecticides. To enhance its genetic tractability, we determined the utility of a ~2 kb An. stephensi lipophorin (lp) promoter fragment in driving transgene expression. Lipophorin genes are involved in lipid transport in insects, and an orthologous promoter in An. gambiae (AGAP001826) was previously demonstrated to successfully express a transgene. In the present study, we qualitatively characterised the expression of a ZsYellow fluorescent marker protein, expressed by An. stephensi lp promoter fragment. Our study indicated that the lp promoter fragment was effective, generating a distinct expression pattern in comparison to the commonly utilised 3xP3 promoter. The lp:ZsYellow fluorescence was largely visible in early instar larvae and appeared more intense in later instar larvae, pupae and adults, becoming especially conspicuous in adult females after a blood meal. Different isolines showed some variation in expression pattern and intensity. Aside from general transgene expression, as the lp promoter produces a suitable fluorescent protein marker expression pattern, it may facilitate genotypic screening and aid the development of more complex genetic biocontrol systems, such as multi-component gene drives. This study represents an expansion of the An. stephensi genetic toolbox, an important endeavour to increase the speed of An. stephensi research and reach public health milestones in combating malaria.

3.
Parasitology ; 151(2): 213-219, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38105582

RESUMO

In 2009, a large outbreak of leishmaniasis, associated with environmental changes, was declared near Madrid (Spain), in which Phlebotomus perniciosus was the vector, whereas the main reservoirs were hares and rabbits. Analysis of isolates from humans, vectors and leporids from the focus identified the Leishmania infantum ITS-Lombardi genotype. However, multilocus enzyme electrophoresis (MLEE), the reference technique for Leishmania typing, and sequencing of the hsp70 gene, a commonly used marker, were not performed. In the present study, 19 isolates from P. perniciosus (n = 11), hares (n = 5) and rabbits (n = 3) from the outbreak area, all characterized as ITS-Lombardi in previous studies, were analysed by MLEE and hsp70 sequencing. The hsp70 results confirmed that all the analysed strains are L. infantum. However, by MLEE, 4 different zymodemes of L. infantum were identified based on variable mobilities of the NP1 enzyme: MON-34 (NP1100, n = 11), MON-80 (NP1130, n = 6), MON-24 (NP1140, n = 1) and MON-331 (NP1150, n = 1). The relative frequency of these zymodemes does not correspond to their usual occurrence in Spain. Moreover, MON-34 and MON-80 were found in P. perniciosus, hares and rabbits for the first time. These findings continue to provide insights into the outbreak and call for further studies with a higher number of strains.


Assuntos
Lebres , Lagomorpha , Leishmania infantum , Humanos , Animais , Coelhos , Espanha/epidemiologia , Leishmania infantum/genética , Surtos de Doenças , Proteínas de Choque Térmico HSP70/genética
4.
Semin Cell Dev Biol ; 72: 10-18, 2017 12.
Artigo em Inglês | MEDLINE | ID: mdl-29127045

RESUMO

The Myogenic Regulatory Factors (MRFs) Myf5, MyoD, myogenin and MRF4 are members of the basic helix-loop-helix family of transcription factors that control the determination and differentiation of skeletal muscle cells during embryogenesis and postnatal myogenesis. The dynamics of their temporal and spatial expression as well as their biochemical properties have allowed the identification of a precise and hierarchical relationship between the four MRFs. This relationship establishes the myogenic lineage as well as the maintenance of the terminal myogenic phenotype. The application of genome-wide technologies has provided important new information as to how the MRFs function to activate muscle gene expression. Application of combined functional genomics technologies along with single cell lineage tracing strategies will allow a deeper understanding of the mechanisms mediating myogenic determination, cell differentiation and muscle regeneration.


Assuntos
Diferenciação Celular/genética , Linhagem da Célula/genética , Desenvolvimento Muscular/genética , Músculo Esquelético/metabolismo , Fatores de Regulação Miogênica/genética , Regeneração/genética , Animais , Regulação da Expressão Gênica no Desenvolvimento , Camundongos , Músculo Esquelético/citologia , Músculo Esquelético/embriologia , Fatores de Regulação Miogênica/classificação , Filogenia
5.
Cell Mol Life Sci ; 73(15): 2897-910, 2016 08.
Artigo em Inglês | MEDLINE | ID: mdl-27026300

RESUMO

During organism development, a diversity of cell types emerges with disparate, yet stable profiles of gene expression with distinctive cellular functions. In addition to gene promoters, the genome contains enhancer regulatory sequences, which are implicated in cellular specialization by facilitating cell-type and tissue-specific gene expression. Enhancers are DNA binding elements characterized by highly sophisticated and various mechanisms of action allowing for the specific interaction of general and tissue-specific transcription factors (TFs). However, eukaryotic organisms package their genetic material into chromatin, generating a physical barrier for TFs to interact with their cognate sequences. The ability of TFs to bind DNA regulatory elements is also modulated by changes in the chromatin structure, including histone modifications, histone variants, ATP-dependent chromatin remodeling, and the methylation status of DNA. Furthermore, it has recently been revealed that enhancer sequences are also transcribed into a set of enhancer RNAs with regulatory potential. These interdependent processes act in the context of a complex network of chromatin interactions, which together contributes to a renewed vision of how gene activation is coordinated in a cell-type-dependent manner. In this review, we describe the interplay between genetic and epigenetic aspects associated with enhancers and discuss their possible roles on enhancer function.


Assuntos
Montagem e Desmontagem da Cromatina , Elementos Facilitadores Genéticos , Animais , Metilação de DNA , Epigênese Genética , Código das Histonas , Humanos , Regiões Promotoras Genéticas , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Ativação Transcricional
6.
Biochim Biophys Acta ; 1839(11): 1233-41, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25239823

RESUMO

Gene promoters are frequently insufficient to drive the spatiotemporal patterns of gene expression during cell differentiation and organism development. Enhancers convey these properties through diverse mechanisms, including long-distance interactions with target promoters via their association with specific transcription factors. Despite unprecedented progress in the knowledge of enhancer mechanisms of action, there are still many unanswered questions. In particular, the contribution of an enhancer's local chromatin configuration to its mechanism of action is not completely understood. Here we describe a novel regulatory element, the Upstream Enhancer Element (UEE), which modulates the activity of the chicken α-globin 3' enhancer by regulating its chromatin structure, specifically by positioning a nucleosome upstream of the core enhancer. This element binds nuclear factors and confers a more restricted activation on the α-globin 3' enhancer, suggesting a progressive/rheostatic model for enhancer activity. Our results suggest that the UEE activity contributes to the positioning of a nucleosome that is necessary for the α-globin 3' enhancer activation.


Assuntos
Regiões 3' não Traduzidas , Galinhas/genética , Cromatina/química , Elementos Facilitadores Genéticos , alfa-Globinas/genética , Animais , Embrião de Galinha , Cromatina/metabolismo , Conformação de Ácido Nucleico , Nucleossomos/metabolismo , Transcrição Gênica
7.
Vet Res ; 46: 138, 2015 Dec 09.
Artigo em Inglês | MEDLINE | ID: mdl-26645907

RESUMO

A human leishmaniasis outbreak is occurring in the Madrid region, Spain, with the parasite and vector involved being Leishmania infantum and Phlebotomus perniciosus respectively. The aim of this study was to investigate the virulence of L. infantum isolates from the focus using a natural transmission model. Hamsters were infected by intraperitoneal inoculation (IP) or by bites of sand flies experimentally infected with L. infantum isolates obtained from P. perniciosus collected in the outbreak area (IPER/ES/2012/BOS1FL1 and IPER/ES/2012/POL2FL6) and a well characterized L. infantum strain JPCM5 (MCAN/ES/98/LLM-877). Hamster infections were monitored by clinical examination, serology, culture, parasite burden, Giemsa-stained imprints, PCR, histopathology and xenodiagnostic studies. Establishment of infection of L. infantum was achieved with the JPCM5 strain and outbreak isolates by both P. perniciosus infective bites or IP route. However, high virulence of BOS1FL1 and POL2FL6 isolates was highlighted by the clinical outcome of disease, high parasite detection in spleen and liver, high parasitic loads and positivity of Leishmania serology. Transmission by bite of POL2FL6 infected flies generated a slower progression of clinical disease than IP infection, but both groups were infective to P. perniciosus by xenodiagnosis at 2 months post-infection. Conversely, hamsters inoculated with JPCM5 were not infective to sand flies. Histopathology studies confirmed the wide spread of POL2FL6 parasites to several organs. A visceral leishmaniasis model that mimics the natural transmission in nature allowed us to highlight the high virulence of isolates that are circulating in the focus. These findings contribute to a better understanding of the outbreak epidemiology.


Assuntos
Leishmania infantum/fisiologia , Leishmania infantum/patogenicidade , Leishmaniose Visceral/transmissão , Phlebotomus/parasitologia , Animais , Feminino , Humanos , Masculino , Mesocricetus , Espanha , Virulência
8.
Front Aging Neurosci ; 16: 1420072, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39026994

RESUMO

Introduction: Studies on the aging brain often occur in active settings, but comparatively few investigate brain activity in resting states. However, exploring brain activity in a resting state offers valuable insights into spontaneous neural processes unaffected by task-specific influences. Objective: To investigate the relationship between self-care practices, cognitive function, and patterns of brain activity in healthy older adults, taking into account predictions from aging brain models. Methodology: 77 older adults aged 61 to 87 completing a self-care practices questionnaire, neuropsychological tests, and resting-state electroencephalogram (EEG) recordings. Participants were classified into two groups according to their self-care practices: traditional self-care (T-SC) and developmental self-care (D-SC). Results: Although neuropsychological tests did not yield significant differences between the D-SC and T-SC groups, patterns of brain activity revealed distinct behaviors. The T-SC group demonstrated patterns more consistent with established aging brain models, contrasting with the D-SC group, which exhibited brain activity akin to that observed in younger adults. Specifically, the T-SC group displayed hyperactivation related to memory and executive function performance, alongside heightened alpha power in posterior regions. Furthermore, bilateral frontal activation in the beta band was evident. Conclusions: The findings suggest a nuanced relationship between self-care practices and brain activity in older adults. While the T-SC group demonstrated brain activity patterns consistent with conservative aging, indicating the preservation of typical aging characteristics, the D-SC group displayed activity suggestive of a potential protective effect. This effect may be linked to self-care strategies that foster development and resilience in cognitive aging.

9.
Ticks Tick Borne Dis ; 15(6): 102401, 2024 Oct 08.
Artigo em Inglês | MEDLINE | ID: mdl-39383837

RESUMO

Red foxes (Vulpes vulpes) are found throughout the United Kingdom (UK), and can reach high population densities in urban areas. They are often infested with ticks which may carry tick-borne pathogens, leading to a risk of transmission to domestic animals and humans. This study investigated the prevalence of tick-borne pathogens in ticks sourced from red fox carcasses across Great Britain between 2018 and 2022. Tick species were identified using morphological keys and molecular barcoding, followed by specific pathogen testing using PCR. In total, 227 ticks were collected from 93 foxes. Pooling (n = 2) was undertaken for unengorged nymphs from the same tick species and fox host, with 203 homogenates tested in total (24 pools and 179 individual ticks). Ixodes hexagonus was the most abundant tick species sampled (73 %), of which 59 % were nymphs and 41 % were females. Less common were Ixodes ricinus (12 %) and Ixodes canisuga (15 %), the majority of which were females (73 % and 91 %, respectively). One Ixodes sp. larva was identified. Babesia DNA was identified in seven individual ticks and once in pooled ticks (n = 2); seven detections were in I. hexagonus and one in I. canisuga, with an overall detection rate of 7 % (95 % CI: 6 - 8 %). Sequence analysis confirmed that all Babesia detections in I. hexagonus were Babesia vulpes, with detection of Babesia Badger Type A in I. canisuga. Screening for Anaplasma phagocytophilum DNA through amplification of the msp2 gene yielded an overall detection rate of 4 % (detected in I. hexagonus only). Louping ill virus was not detected by qRT-PCR in any tick RNA tested. The majority of pathogen detections were in ticks from red foxes in rural areas of the UK, although a small number of Babesia detections were in ticks collected from semi-rural or urban red foxes. Additionally, B. vulpes was detected in GB red fox tissues, suggesting a potential role as a reservoir host. This study confirms the detection of tick-borne pathogens in ticks infesting UK red foxes and highlights the involvement of GB tick species in animal or human disease transmission.

10.
Nat Commun ; 15(1): 729, 2024 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-38272895

RESUMO

Aedes aegypti is the main vector of several major pathogens including dengue, Zika and chikungunya viruses. Classical mosquito control strategies utilizing insecticides are threatened by rising resistance. This has stimulated interest in new genetic systems such as gene drivesHere, we test the regulatory sequences from the Ae. aegypti benign gonial cell neoplasm (bgcn) homolog to express Cas9 and a separate multiplexing sgRNA-expressing cassette inserted into the Ae. aegypti kynurenine 3-monooxygenase (kmo) gene. When combined, these two elements provide highly effective germline cutting at the kmo locus and act as a gene drive. Our target genetic element drives through a cage trial population such that carrier frequency of the element increases from 50% to up to 89% of the population despite significant fitness costs to kmo insertions. Deep sequencing suggests that the multiplexing design could mitigate resistance allele formation in our gene drive system.


Assuntos
Aedes , Tecnologia de Impulso Genético , Inseticidas , Infecção por Zika virus , Zika virus , Animais , Sistemas CRISPR-Cas/genética , Aedes/genética , RNA Guia de Sistemas CRISPR-Cas , Infecção por Zika virus/genética , Zika virus/genética
11.
Parasitol Res ; 112(7): 2453-9, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23535889

RESUMO

Since 2010, the number of cases of both human visceral leishmaniasis and cutaneous leishmaniasis in southwestern Madrid region (Spain) and more specifically in the town of Fuenlabrada has increased. Direct xenodiagnosis of leishmaniasis proved that hares (Lepus granatensis) from this focus are able to infect with Leishmania infantum colonized Phlebotomus perniciosus. To a better understanding of this focus of leishmaniasis, we conducted an entomological survey using CDC light traps, at the end of the seasonal transmission period of 2011 before the beginning of control measures of the disease, to study the phlebotomine sand flies species involved. Detection of Leishmania DNA in the sand flies captured was studied by kDNA-PCR and cpb-PCR. In addition, blood fed and gravid female P. perniciosus were analysed by a PCR based in vertebrate cytochrome b (cyt b) gene. Taxonomic identification of captured sand flies (n = 174) as P. perniciosus (n = 171) and Sergentomyia minuta (n = 3) together with the analysis of blood feeding in ten sand flies that shows a high preference for hares (n = 6), followed by humans (n = 3), and cats (n = 1) confirm a strong association between P. perniciosus hares and humans in the focus. Moreover, 79 out of 135 (58.5 %) P. perniciosus were positive to L. infantum by PCR approaches. These data support the increase of human leishmaniasis cases in the area and the existence of an unusual sylvatic cycle alternative to the classical domestic one, where the dog is the main reservoir of L. infantum.


Assuntos
Sangue/parasitologia , Comportamento Alimentar , Leishmania infantum/isolamento & purificação , Phlebotomus/fisiologia , Phlebotomus/parasitologia , Animais , Gatos , Cães , Entomologia/métodos , Humanos , Leishmaniose/epidemiologia , Parasitologia/métodos , Phlebotomus/classificação , Reação em Cadeia da Polimerase/métodos , Coelhos , Espanha/epidemiologia
12.
Front Public Health ; 11: 1254172, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37876713

RESUMO

Introduction: The current notion of "care in old age" should be reconceptualized in the ageing societies of the 21st century. Currently, "being old" means that one is actively involved in their care and has the desire to retain control and independence. Objective: Understand and analyze the efficacy of interventions in the physical and psychological self-care practices of healthy community-dwelling older people. Methodology: Systematic review and meta-analysis. The guidelines of the PRISMA guide were followed. The methodological quality of the studies was checked using Cochrane Effective Practice and Organisation of Care criteria, and the search was performed between 2016 and 2021. Results: Of the 1,866 evaluated, 8 studies met the criteria. The systematic review reveals that self-care interventions focus on physical health-related variables but not on psychological variables. The meta-analysis shows that interventions significantly improve physical health-related variables (care visits, hospital admission, medication, and gait speed). Conclusion: Self-care training programs should include psychological variables to increase health and well-being in healthy older people.


Assuntos
Vida Independente , Autocuidado , Humanos , Idoso , Hospitalização , Nível de Saúde
13.
Geriatrics (Basel) ; 8(3)2023 May 13.
Artigo em Inglês | MEDLINE | ID: mdl-37218834

RESUMO

Two competing psychological approaches for how to care for oneself to stay healthy in old age have coexisted and dominated the scientific literature. OBJECTIVE: Identify the self-care practices of healthy older adults and establish the relationship between these practices and the cognitive processes involved. METHOD: 105 healthy older people (83.91% women) recorded their self-care practices using the Care Time Test and underwent a cognitive evaluation. RESULTS: The frequency and variety of different activities that participants spent performing on a day of the week where they had the fewest obligations are as follows: nearly 7 h on seven survival activities, 4 h and 30 min on three maintenance of functional independence activities and 1 h on one activity that promoted personal development. Older people who carry out activities in a developmental approach showed better everyday memory (8.63 points) and attention levels (7.00 points) than older people who carry out activities using a conservative approach (memory: 7.43; attention level: 6.40). CONCLUSION: The results evidenced that the frequency and variety of activities that promote personal development are associated with better attention and memory performance.

14.
Healthcare (Basel) ; 11(8)2023 Apr 13.
Artigo em Inglês | MEDLINE | ID: mdl-37107947

RESUMO

(1) Background: Coping includes the specific cognitive processes and behaviours that the patient uses when faced with the stress of living with a chronic disease. Self-efficacy is the knowledge that individuals have about their abilities and their confidence to face a problem or cope with a situation (disease). The aim of this study was to explore the role of coping and self-efficacy in inflammatory bowel disease. (2) Materials and Methods: A total of 92 participants were included (33 had been diagnosed with Crohn's disease, 23 with ulcerative colitis and 36 were healthy participants). The Coping Strategies Inventory was used to measure which coping strategies were employed, differentiating them as active or passive. The General Self-Efficacy Scale was used to measure self-efficacy. (3) Results: The results indicate that people with inflammatory bowel disease used strategies related to passive coping more than healthy people (mean of 36.39 ± 13.92 vs. 29.77 ± 10.70, p = 0.017). Additionally, people with inflammatory bowel disease used social withdrawal more than healthy participants (mean of 8.30 ± 5.07 vs. 4.47 ± 4.17, p < 0.001). In addition, there are significant differences in emotion-focused engagement coping strategies. People with inflammatory bowel disease used this strategy less than healthy people (mean of 21.77 ± 7.75 vs. 25.03 ± 7.00, p = 0.044). Finally, healthy participants used the emotion-focused disengagement strategy less than those diagnosed with inflammatory bowel disease (mean of 9.81 ± 7.74 vs. 15.61 ± 10.14, p = 0.004). (4) Conclusions: Actions aimed at the development of active coping strategies and patient socialisation must be included in the treatment of inflammatory bowel disease.

15.
Medicina (B Aires) ; 83(6): 918-926, 2023.
Artigo em Espanhol | MEDLINE | ID: mdl-38117711

RESUMO

INTRODUCTION: The second victims are those professionals who, when faced with a mistake, are traumatized, questioning their professional and personal skills. Knowledge of their experience allows institutions to understand the consequences of error in professionals to carry out care actions for the professionals involved, improving care security. The objective of this study is to characterize the phenomenon of second victims and to know the perceived support measures. METHOD: Cross-sectional descriptive observational quantitative study. Health professionals from five highly complex institutions participated through convenience sampling. The data collection instrument was the Second Victim Experience and Support Tool (SVEST) validated in Argentina, composed of impact dimensions (psychological, physical and professional skills) of support received (colleagues, supervisors, institutional and measures not related to the workplace) and two outcome dimensions (professional abandonment and absenteeism). RESULTS: 1134 professionals participated and 56% said they had made a mistake. The mean STEVS was 2.5, with the psychological impact dimension predominating (x = 3.4). As part of the support measures, professionals perceived greater support from family and friends (not related to work) (x = 3.1). The greater the impact, the lower the perception of support measures (Rho: -0.8, p = 0.047). DISCUSSION: More than half of the professionals refer to being second victim with a significant psychological impact, who, despite working in institutions that have patient safety protocols, received support from nonwork-related environments.


Introducción: Las segundas víctimas son aquellos profesionales que, ante un error, quedan traumatizados, cuestionándose sus habilidades profesionales y personales. El conocimiento de su experiencia permite a las instituciones entender las consecuencias del error en los profesionales para realizar acciones de atención a los profesionales involucrados mejorando la seguridad de atención. El objetivo de este estudio es caracterizar el fenómeno de segundas víctimas y conocer las medidas de apoyo percibidas. Métodos: Estudio cuantitativo observacional descriptivo transversal. Participaron profesionales de la salud de cinco instituciones de alta complejidad a través de un muestreo por conveniencia. El instrumento de recogida de datos fue el Second Victim Experience and Support Tool (SVEST) validado en Argentina, compuesta por dimensiones de impacto (psicológico, físico y de habilidades profesionales) de apoyo recibido (colegas, supervisores, institucional y medidas no relacionadas al ámbito laboral) y dos dimensiones de resultado (abandono profesional y ausentismo). Resultados: Participaron 1134 profesionales. El 56% manifestó haber cometido un error y completó el SVEST. La media del SVEST fue de 2.5 predominando la dimensión de impacto psicológico (x = 3.4). Como parte de las medidas de apoyo los profesionales percibieron mayor apoyo en familiares y amigos (x = 3.1). A mayor impacto menor percepción de las medidas de apoyo (Rho: -0.8, p = 0.047). Discusión: Más de la mitad de los profesionales refieren ser segundas víctimas con un impacto psicológico importante, los cuales, a pesar de trabajar en instituciones que cuentan con protocolos de seguridad del paciente, reciben el apoyo principalmente de entornos no relacionados al trabajo.


Assuntos
Pessoal de Saúde , Local de Trabalho , Humanos , Argentina , Estudos Transversais , Pessoal de Saúde/psicologia , Segurança do Paciente , Inquéritos e Questionários
16.
Nat Commun ; 14(1): 338, 2023 01 20.
Artigo em Inglês | MEDLINE | ID: mdl-36670107

RESUMO

CRISPR/Cas9-based homing gene drives have emerged as a potential new approach to mosquito control. While attempts have been made to develop such systems in Aedes aegypti, none have been able to match the high drive efficiency observed in Anopheles species. Here we generate Ae. aegypti transgenic lines expressing Cas9 using germline-specific regulatory elements and assess their ability to bias inheritance of an sgRNA-expressing element (kmosgRNAs). Four shu-Cas9 and one sds3-Cas9 isolines can significantly bias the inheritance of kmosgRNAs, with sds3G1-Cas9 causing the highest average inheritance of ~86% and ~94% from males and females carrying both elements outcrossed to wild-type, respectively. Our mathematical model demonstrates that sds3G1-Cas9 could enable the spread of the kmosgRNAs element to either reach a higher (by ~15 percentage point) maximum carrier frequency or to achieve similar maximum carrier frequency faster (by 12 generations) when compared to two other established split drive systems.


Assuntos
Aedes , Tecnologia de Impulso Genético , Animais , Masculino , Feminino , Aedes/genética , Animais Geneticamente Modificados , Sequências Reguladoras de Ácido Nucleico
17.
J Phys Condens Matter ; 34(24)2022 Apr 14.
Artigo em Inglês | MEDLINE | ID: mdl-35358960

RESUMO

We investigated by first principle calculations the adsorption of Liq(q= -1, 0 or +1) on a silicene single layer. Pristine and three different defective silicene configurations with and without Li doping were studied: single vacancy (SV), double vacancy (DV) and Stone-Wales (STW). Structural studies and the adsorption energies of various sites were obtained and compared in order to understand the stability of the Li on the surface. Moreover, electronic structure and charge density difference analysis were performed before and after adsorption at the most stables sites, which showed the presence of a magnetic moment in the undoped SV system, the displacement of the Fermi level produced by Li doping and a charge transfer from Li to the surface. Additionally, quantum capacity (QC) and charge density studies were performed on these systems. This analysis showed that the generation of defects and doping improves the QC of silicene in positive bias, because of the existence of 3p orbital in the zone of the defect. Consequently, the innovative calculations performed in this work of charged lithium doping on silicene can be used for future comparison with experimental studies of this Li-ion battery anode material candidate.

18.
Nat Commun ; 13(1): 7145, 2022 11 21.
Artigo em Inglês | MEDLINE | ID: mdl-36414618

RESUMO

CRISPR/Cas gene drives can bias transgene inheritance through different mechanisms. Homing drives are designed to replace a wild-type allele with a copy of a drive element on the homologous chromosome. In Aedes aegypti, the sex-determining locus is closely linked to the white gene, which was previously used as a target for a homing drive element (wGDe). Here, through an analysis using this linkage we show that in males inheritance bias of wGDe did not occur by homing, rather through increased propagation of the donor drive element. We test the same wGDe drive element with transgenes expressing Cas9 with germline regulatory elements sds3, bgcn, and nup50. We only find inheritance bias through homing, even with the identical nup50-Cas9 transgene. We propose that DNA repair outcomes may be more context dependent than anticipated and that other previously reported homing drives may, in fact, bias their inheritance through other mechanisms.


Assuntos
Aedes , Tecnologia de Impulso Genético , Masculino , Sistemas CRISPR-Cas/genética , Endonucleases/genética , Células Germinativas , Padrões de Herança/genética , Aedes/genética , Animais , Transgenes
19.
PLoS Negl Trop Dis ; 15(3): e0009240, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33720936

RESUMO

BACKGROUND: An outbreak of human leishmaniasis due to Leishmania infantum has been registered in an urban area of southwestern Madrid, Spain, since 2010. Entomological surveys carried out in the municipalities of Fuenlabrada, Leganés, Getafe and Humanes de Madrid showed that Phlebotomus perniciosus is the only potential vector. In this work, an intensive molecular surveillance was performed in P. perniciosus females captured in the region between 2012 and 2018. METHODOLOGY/PRINCIPAL FINDINGS: A total of 1805 P. perniciosus females were analyzed for Leishmania infection, and 1189 of them also for bloodmeal identification. Eleven different species of vertebrate were detected by amplification and subsequent sequencing of the 359 bp cytb fragment. The most prevalent blood source identified was hare (n = 553, 46.51%), followed by rabbit (n = 262, 21.95%). Less frequent were cat (n = 45, 3.80%), human (n = 34, 2.90%), pig (n = 14, 1.20%), horse (n = 11, 0.93%), sheep (n = 3, 0.25%), rhea (n = 3, 0.25%), partridge (n = 1, 0.09%) and chicken (n = 1, 0.09%). The distribution of the blood meal sources varied between the different locations. Regarding L. infantum detection, PCR amplification of a fragment of kDNA, cpb gene and ITS1 region showed 162 positive specimens (8.97%). The highest infection rate was found in the municipality of Leganés (15.17%). CONCLUSIONS: The results of this molecular survey in P. perniciosus, the only leishmaniasis vector in the outbreak occurred in southwestern Madrid region, showed its opportunistic blood-feeding behaviour, high infection rates and the differences between the different points. This study was an essential part of the intensive surveillance plan in the area and the results obtained have supported the implementation of control measures in the outbreak.


Assuntos
Comportamento Alimentar , Leishmania infantum/isolamento & purificação , Leishmaniose Visceral/epidemiologia , Phlebotomus/parasitologia , Animais , Surtos de Doenças , Feminino , Humanos , Leishmaniose Visceral/prevenção & controle , Leishmaniose Visceral/transmissão , Espanha/epidemiologia
20.
Artigo em Inglês | MEDLINE | ID: mdl-33182788

RESUMO

Background: The literature favors discussion on socio-spatial conditions at the macro- (city) and micro- (housing) level that promote healthy aging in place. Objectives: (a) Identify the association between physical and social characteristics of the family home and the functional level and quality of life of older people and (b) provide normative data on adequate/inadequate households based on the Home Observation for Measurement of the Environment (HOME) inventory and the Spanish Informant Questionnaire on Cognitive Decline in the Elderly (S-IQCODE) test. Methods: In total, 79 healthy older adults completed the HOME inventory and the Montreal Cognitive Assessment (MoCA), S-IQCODE, Kessler Psychological Distress Scale (K-10) and ICEpop CAPability measure for Older people (ICECAP-O) tests. A regression model, the effect size and the means of the scores of HOME (adequate/inadequate) test and the cognitive level (optimal/normal) were calculated. Results: The regression model discloses that adequate home scores are associated with cognitive level (odds ratio (OR): 0.955, confidence interval (CI)95%: 0.918-0.955); quality of life (OR: 6.542, CI95%: 1.750-24.457), living with other people (OR: 5.753, CI95%: 1.456-22.733) and level of education (OR: 0.252, CI95%: 0.064-0.991). The normative data between HOME and S-IQCODE scores showed a good adjustment (d = 0.70). Conclusion: There is a significant relationship between the physical environment of the home and personal variables (sociodemographic information, quality of life and cognitive functionality). In addition, from this last variable, the normative data of an adequate/inadequate household for an older person have been established.


Assuntos
Habitação para Idosos , Vida Independente , Qualidade de Vida , Idoso , Idoso de 80 Anos ou mais , Feminino , Habitação para Idosos/normas , Humanos , Vida Independente/normas , Masculino , Testes de Estado Mental e Demência , Espanha/epidemiologia , Inquéritos e Questionários
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