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INTRODUCTION: With the emergence of newer SARS-CoV-2 variants and their substantial effects on the levels and duration of protection against infection, an understanding of these characteristics of the protection conferred by humoral and cellular immunity can aid in the proper development and implementation of vaccine and safety guidelines. METHODS: We conducted a rapid literature review and searched five electronic databases weekly from 1 November 2021 to 30 September 2022. Studies that assessed the humoral or cellular immunity conferred by infection, vaccination or a hybrid (combination of both) in adults and risk groups (immunocompromised and older populations) were identified. Studies were eligible when they reported data on immunological assays of COVID-19 (related to vaccination and/or infection) or the effectiveness of protection (related to the effectiveness of vaccination and/or infection). RESULTS: We screened 5103 studies and included 205 studies, of which 70 provided data on the duration of protection against SARS-CoV-2 infection. The duration of protection of adaptive immunity was greatly impacted by Omicron and its subvariants: levels of protection were low by 3-6 months from exposure to infection/vaccination. Although more durable, cellular immunity also showed signs of waning by 6 months. First and second mRNA vaccine booster doses increased the levels of protection against infection and severe disease from Omicron and its subvariants but continued to demonstrate a high degree of waning over time. CONCLUSION: All humoral immunities (infection-acquired, vaccine-acquired and hybrid) waned by 3-6 months. Cellular immunity was more durable but showed signs of waning by 6 months. Hybrid immunity had the highest magnitude of protection against SARS-CoV-2 infection. Boosting may be recommended as early as 3-4 months after the last dose, especially in risk groups.
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Vacunas contra la COVID-19 , COVID-19 , Inmunidad Celular , SARS-CoV-2 , Humanos , COVID-19/prevención & control , COVID-19/inmunología , SARS-CoV-2/inmunología , Vacunas contra la COVID-19/inmunología , Inmunidad Celular/inmunología , Inmunidad Humoral/inmunología , Inmunización Secundaria , VacunaciónRESUMEN
This study conducts a comprehensive analysis of forensic toxicology research trends, publication patterns, author's contributions, and collaboration. Utilizing the Scopus database, we scrutinized 3259 articles across 348 journals spanning from 1975 to 2023. Analysis employed diverse software tools such as VOSviewer, RStudio, MS Excel, and MS Access to dissect various publication aspects. We observed a notable surge in publications post-2007, indicating heightened research interest. Leading contributors included the United States, Germany, and Italy, with Logan B.K. emerging as the most prolific author. Forensic Science International stood out as the primary journal, publishing 888 articles and accruing significant citations. Keyword co-occurrences such as "forensic toxicology," "forensic science," and "toxicology" underscored core thematic areas in the field. Moreover, extensive research collaboration, especially among Western nations in Europe, was evident. This study underscores the imperative for enhanced collaboration between developing and developed nations to foster further advancements in forensic science. Strengthened partnerships can catalyze innovation, facilitate knowledge dissemination, and address emerging challenges, thereby propelling the field of forensic toxicology toward new frontiers of discovery and application.
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Toxicología Forense , Toxicología Forense/tendencias , Humanos , Bibliometría , Publicaciones Periódicas como Asunto/tendencias , Publicaciones Periódicas como Asunto/estadística & datos numéricos , Investigación Biomédica/tendencias , Edición/tendencias , Edición/estadística & datos numéricosRESUMEN
BACKGROUND: Guaranteeing durability, provenance, accessibility, and trust in open data sets can be challenging for researchers and organizations that rely on public repositories of data critical for epidemiology and other health analytics. The required data repositories are often difficult to locate and may require conversion to a standard data format. Data-hosting websites may also change or become unavailable without warning. A single change to the rules in one repository can hinder updating a public dashboard reliant on data pulled from external sources. These concerns are particularly challenging at the international level, because policies on systems aimed at harmonizing health and related data are typically dictated by national governments to serve their individual needs. OBJECTIVE: In this paper, we introduce a comprehensive public health data platform, EpiGraphHub, that aims to provide a single interoperable repository for open health and related data. METHODS: The platform, curated by the international research community, allows secure local integration of sensitive data while facilitating the development of data-driven applications and reports for decision-makers. Its main components include centrally managed databases with fine-grained access control to data, fully automated and documented data collection and transformation, and a powerful web-based data exploration and visualization tool. RESULTS: EpiGraphHub is already being used for hosting a growing collection of open data sets and for automating epidemiological analyses based on them. The project has also released an open-source software library with the analytical methods used in the platform. CONCLUSIONS: The platform is fully open source and open to external users. It is in active development with the goal of maximizing its value for large-scale public health studies.
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Análisis de Datos , Salud Pública , Humanos , Recolección de Datos , Bases de Datos Factuales , Gobierno FederalRESUMEN
Investigating drowning-related deaths remains a significant problem for forensic personnel all over the world. The previously published decomposition scoring method like the total aquatic decomposition (TAD) score promises to estimate the correct post-mortem submersion interval (PMSI) in aquatic habitats through the assessment and calculation of the decomposition rate and accumulated degree days (ADD). The current study comprised of 53 drowned death cases belonging to various districts of Haryana from May 2016 to August 2017. The regression and Pearson's correlation indicated a significant correlation between the TAD scores and the actual ADD (calculated through water temperatures) (r2 = 0.917) and between the actual and the estimated ADDs (calculated through TAD scores used by Heaton et al. [21]) (r = 0.9585). The results indicated that the estimated ADD tends to over predict the PMSI compared to the actual ADD. It is further confirmed by paired t-test, which showed the mean of actual ADD (mean = 349) to be significantly lower than the mean of estimated ADD (mean = 663). Moreover, these methods will help forensic investigators and researchers formulate region-specific regression equations for PMSI estimation.
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Ahogamiento , Humanos , Cambios Post Mortem , Inmersión , Cadáver , Autopsia , Patologia Forense/métodosRESUMEN
Chrysomya megacephala and C. rufifacies are globally predominant species of blowflies that are amongst the primary colonisers of decomposing carcasses. The development of blowflies has been studied extensively to estimate the minimum postmortem interval (PMImin). This article evaluates the effect of aluminium phosphide (AlP) on the larval morphometry and life cycles of C. megacephala and C. rufifacies. The rabbits were given different dosages of AlP and their livers were excised to rear C. megacephala and C. rufifacies. Through multiple-regression analysis, we assessed the overall impact of varying dosages of AlP on larvae of both species. AlP accelerated development until pupation, whilst the time until emergence remained the same. At higher concentrations, AlP had a positive effect on the development of these species. Since both species are abundant, forensic entomology studies examining the effect of AlP on morphometry and developmental stages would provide valuable information to investigators assessing PMImin.
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Emerging infectious diseases are a growing threat in sub-Saharan African countries, but the human and technical capacity to quickly respond to outbreaks remains limited. Here, we describe the experience and lessons learned from a joint project with the WHO Regional Office for Africa (WHO AFRO) to support the sub-Saharan African COVID-19 response.In June 2020, WHO AFRO contracted a number of consultants to reinforce the COVID-19 response in member states by providing actionable epidemiological analysis. Given the urgency of the situation and the magnitude of work required, we recruited a worldwide network of field experts, academics and students in the areas of public health, data science and social science to support the effort. Most analyses were performed on a merged line list of COVID-19 cases using a reverse engineering model (line listing built using data extracted from national situation reports shared by countries with the Regional Office for Africa as per the IHR (2005) obligations). The data analysis platform The Renku Project ( https://renkulab.io ) provided secure data storage and permitted collaborative coding.Over a period of 6 months, 63 contributors from 32 nations (including 17 African countries) participated in the project. A total of 45 in-depth country-specific epidemiological reports and data quality reports were prepared for 28 countries. Spatial transmission and mortality risk indices were developed for 23 countries. Text and video-based training modules were developed to integrate and mentor new members. The team also began to develop EpiGraph Hub, a web application that automates the generation of reports similar to those we created, and includes more advanced data analyses features (e.g. mathematical models, geospatial analyses) to deliver real-time, actionable results to decision-makers.Within a short period, we implemented a global collaborative approach to health data management and analyses to advance national responses to health emergencies and outbreaks. The interdisciplinary team, the hands-on training and mentoring, and the participation of local researchers were key to the success of this initiative.
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COVID-19 , África del Sur del Sahara/epidemiología , COVID-19/epidemiología , Brotes de Enfermedades/prevención & control , Humanos , Salud Pública , Recursos HumanosRESUMEN
During the first wave of the COVID-19 pandemic, sub-Saharan African countries experienced comparatively lower rates of SARS-CoV-2 infections and related deaths than in other parts of the world, the reasons for which remain unclear. Yet, there was also considerable variation between countries. Here, we explored potential drivers of this variation among 46 of the 47 WHO African region Member States in a cross-sectional study. We described five indicators of early COVID-19 spread and severity for each country as of 29 November 2020: delay in detection of the first case, length of the early epidemic growth period, cumulative and peak attack rates and crude case fatality ratio (CFR). We tested the influence of 13 pre-pandemic and pandemic response predictor variables on the country-level variation in the spread and severity indicators using multivariate statistics and regression analysis. We found that wealthier African countries, with larger tourism industries and older populations, had higher peak (p<0.001) and cumulative (p<0.001) attack rates, and lower CFRs (p=0.021). More urbanised countries also had higher attack rates (p<0.001 for both indicators). Countries applying more stringent early control policies experienced greater delay in detection of the first case (p<0.001), but the initial propagation of the virus was slower in relatively wealthy, touristic African countries (p=0.023). Careful and early implementation of strict government policies were likely pivotal to delaying the initial phase of the pandemic, but did not have much impact on other indicators of spread and severity. An over-reliance on disruptive containment measures in more resource-limited contexts is neither effective nor sustainable. We thus urge decision-makers to prioritise the reduction of resource-based health disparities, and surveillance and response capacities in particular, to ensure global resilience against future threats to public health and economic stability.
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COVID-19 , Pandemias , Estudios Transversales , Humanos , SARS-CoV-2 , Organización Mundial de la SaludRESUMEN
The geographic and economic characteristics unique to island nations create a different set of conditions for, and responses to, the spread of a pandemic compared with those of mainland countries. Here, we aimed to describe the initial period of the COVID-19 pandemic, along with the potential conditions and responses affecting variation in the burden of infections and severe disease burden, across the six island nations of the WHO's Africa region: Cabo Verde, Comoros, Madagascar, Mauritius, São Tomé e Príncipe and Seychelles. We analysed the publicly available COVID-19 data on confirmed cases and deaths from the beginning of the pandemic through 29 November 2020. To understand variation in the course of the pandemic in these nations, we explored differences in their economic statuses, healthcare expenditures and facilities, age and sex distributions, leading health risk factors, densities of the overall and urban populations and the main industries in these countries. We also reviewed the non-pharmaceutical response measures implemented nationally. We found that the burden of SARS-CoV-2 infection was reduced by strict early limitations on movement and biased towards nations where detection capacity was higher, while the burden of severe COVID-19 was skewed towards countries that invested less in healthcare and those that had older populations and greater prevalence of key underlying health risk factors. These findings highlight the need for Africa's island nations to invest more in healthcare and in local testing capacity to reduce the need for reliance on border closures that have dire consequences for their economies.
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COVID-19 , Gripe Humana , Atención a la Salud , Humanos , Gripe Humana/epidemiología , Pandemias , SARS-CoV-2RESUMEN
INTRODUCTION: Since sex-based biological and gender factors influence COVID-19 mortality, we wanted to investigate the difference in mortality rates between women and men in sub-Saharan Africa (SSA). METHOD: We included 69 580 cases of COVID-19, stratified by sex (men: n=43 071; women: n=26 509) and age (0-39 years: n=41 682; 40-59 years: n=20 757; 60+ years: n=7141), from 20 member nations of the WHO African region until 1 September 2020. We computed the SSA-specific and country-specific case fatality rates (CFRs) and sex-specific CFR differences across various age groups, using a Bayesian approach. RESULTS: A total of 1656 deaths (2.4% of total cases reported) were reported, with men accounting for 70.5% of total deaths. In SSA, women had a lower CFR than men (mean [Formula: see text] = -0.9%; 95% credible intervals (CIs) -1.1% to -0.6%). The mean CFR estimates increased with age, with the sex-specific CFR differences being significant among those aged 40 years or more (40-59 age group: mean [Formula: see text] = -0.7%; 95% CI -1.1% to -0.2%; 60+ years age group: mean [Formula: see text] = -3.9%; 95% CI -5.3% to -2.4%). At the country level, 7 of the 20 SSA countries reported significantly lower CFRs among women than men overall. Moreover, corresponding to the age-specific datasets, significantly lower CFRs in women than men were observed in the 60+ years age group in seven countries and 40-59 years age group in one country. CONCLUSIONS: Sex and age are important predictors of COVID-19 mortality globally. Countries should prioritise the collection and use of sex-disaggregated data so as to design public health interventions and ensure that policies promote a gender-sensitive public health response.
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COVID-19 , Adolescente , Adulto , África del Sur del Sahara/epidemiología , Teorema de Bayes , Niño , Preescolar , Estudios Transversales , Femenino , Humanos , Lactante , Recién Nacido , Masculino , Persona de Mediana Edad , SARS-CoV-2 , Adulto JovenRESUMEN
INTRODUCTION: Few data on the COVID-19 epidemiological characteristics among the pediatric population in Africa exists. This paper examines the age and sex distribution of the morbidity and mortality rate in children with COVID-19 and compares it to the adult population in 15 Sub-Saharan African countries. METHODS: A merge line listing dataset shared by countries within the Regional Office for Africa was analyzed. Patients diagnosed within 1 March and 1 September 2020 with a confirmed positive RT-PCR test for SARS-CoV-2 were analyzed. Children's data were stratified into three age groups: 0-4 years, 5-11 years, and 12-17 years, while adults were combined. The cumulative incidence of cases, its medians, and 95% confidence intervals were calculated. RESULTS: 9% of the total confirmed cases and 2.4% of the reported deaths were pediatric cases. The 12-17 age group in all 15 countries showed the highest cumulative incidence proportion in children. Adults had a higher case incidence per 100,000 people than children. CONCLUSION: The cases and deaths within the children's population were smaller than the adult population. These differences may reflect biases in COVID-19 testing protocols and reporting implemented by countries, highlighting the need for more extensive investigation and focus on the effects of COVID-19 in children.
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COVID-19 , Adulto , África del Sur del Sahara/epidemiología , Prueba de COVID-19 , Niño , Preescolar , Humanos , Incidencia , Lactante , Recién Nacido , SARS-CoV-2RESUMEN
Forensic entomotoxicology integrates toxicology into forensic entomology to estimate minimum postmortem interval (PMImin) and circumstances of death where toxicants and poisonous substances are the suspected cause of death. Forensic entomotoxicology not only confirms the presence of toxicants in insects feeding off a cadaver but also studies its effect on the bio-morphometry and growth rate of insects. This review article highlights the effects of various toxicants on forensically important species of dipteran flies. It also discusses the parameters that may affect accuracy in estimation of time since death. The bio-physical effects of toxicants (excluding the analytical approach for qualitative detection) would help understand the trends in forensic entomotoxicological research worldwide.
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Dípteros/anatomía & histología , Dípteros/crecimiento & desarrollo , Entomología Forense , Toxicología Forense , Animales , Investigación/tendenciasRESUMEN
Research on studying the decomposition pattern in aquatic habitat is very sparse worldwide. Due to this void, assessment of the postmortem submersion interval (PMSI) of drowned and submerged cadavers is still inaccurate and imprecise. The current work focused on studying the decomposition pattern and insects associated with submerged pig (Sus scrofa) carcasses during various seasons (winter, spring, summer, and rainy) for two years. The total postmortem submersion interval (PMSI) from the fresh stage till the sunken remains stage varied from 25 days for rainy season (624.5 ADD) to 78 days (1890.5 ADD) for winter season. The spring season at 44 days (1067.5 ADD) and summer season at 31 days (763.5 ADD) had moderate PMSI. During this time interval, five stages of decomposition were studied: submerged fresh, early floating, floating decay, advanced floating decay, and sunken remains. A total of 2385 insect specimens were found to be associated with different decomposition stages. The terrestrial activity during the first stage (submerged fresh) was almost nil as the carcasses were submerged. During the early floating stage (2nd stage), various terrestrial insect species visited the carcass and laid their eggs along with the presence of Chironomidae, Coenagrionidae, Lestidae, Aeshnidae families (order Odonata) were also found associated with this stage. In the floating decay (3rd stage), the insects visiting and colonizing the carcass were from Coenagrionidae, Gomphidae, Aeshnidae, Gomphidae, Libellulidae Chironomidae (Order-Odonata) families along with Calliphoridae (order- Diptera). Species from the group of scavenging aquatic beetles (Hydrophilidae), Enochrus esuriens (Walker), Regimbartia attenuata (Fab), Helochares sp. and burrowing water beetles, Canthydrus laetabilis (Family: Noteridae) and some terrestrial species of beetles, i.e. Saprinus sp., Saprinus pensylvenicus and Necrobia rufipus (Family: Cleridae) were also present during the 3rd, 4th and 5th stages. During advanced floating decay (4th stage) and sunken remains (5th stage), many hemipteran species such as Ranatra digitata and Ranatra filiformis (Fab) along with many coleopterans were found visiting the corpse.
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Conducta Alimentaria , Inmersión , Insectos , Cambios Post Mortem , Estaciones del Año , Animales , Entomología Forense , Patologia Forense , Modelos Animales , PorcinosRESUMEN
Ba5Zn4Gd8O21:Tb3+ nanorods; synthesized via solution combustion route; were found to crystallize in the tetragonal (I4/m, 87) crystal system. The UV excitation at 290 nm of all Ba5Zn4Gd8O21:Tb3+ samples yielded the characteristics emission corresponding to 5D4 â 7F6,5,4,3 transitions in Tb3+ activator (used for Judd-Ofelt analysis). A detailed investigation of photoluminescence decay curves and emission spectra of Ba5Zn4Gd8O21:Tb3+ nanorods provided the radiative lifetime (1.0889 ms) and total radiative transition rates from 5D4 state in Tb3+ ion. The electric-dipole radiative transition probabilities (extracted from total radiative rates i.e. electric-dipole + magnetic-dipole) were used to calculate the Judd-Ofelt intensity parameters (Ω2 = 4.76 × 10-20, Ω4 = 2.11 × 10-20 and Ω6 = 2.00 × 10-20 cm2). The very high quantum efficiency of 5D4 state (81%) suggests their potential use in lighting and display devices. Finally, a large magnitude of peak stimulated emission cross section of 5D4 â 7F5 (16.5838 × 10-20 cm2) transition also claims their promising candidature as a good laser material.
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MOTIVATION: The presence of present-day human contaminating DNA fragments is one of the challenges defining ancient DNA (aDNA) research. This is especially relevant to the ancient human DNA field where it is difficult to distinguish endogenous molecules from human contaminants due to their genetic similarity. Recently, with the advent of high-throughput sequencing and new aDNA protocols, hundreds of ancient human genomes have become available. Contamination in those genomes has been measured with computational methods often developed specifically for these empirical studies. Consequently, some of these methods have not been implemented and tested for general use while few are aimed at low-depth nuclear data, a common feature in aDNA datasets. RESULTS: We develop a new X-chromosome-based maximum likelihood method for estimating present-day human contamination in low-depth sequencing data from male individuals. We implement our method for general use, assess its performance under conditions typical of ancient human DNA research, and compare it to previous nuclear data-based methods through extensive simulations. For low-depth data, we show that existing methods can produce unusable estimates or substantially underestimate contamination. In contrast, our method provides accurate estimates for a depth of coverage as low as 0.5× on the X-chromosome when contamination is below 25%. Moreover, our method still yields meaningful estimates in very challenging situations, i.e. when the contaminant and the target come from closely related populations or with increased error rates. With a running time below 5 min, our method is applicable to large scale aDNA genomic studies. AVAILABILITY AND IMPLEMENTATION: The method is implemented in C++ and R and is available in github.com/sapfo/contaminationX and popgen.dk/angsd.
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ADN Antiguo , Secuenciación de Nucleótidos de Alto Rendimiento , Cromosomas , Humanos , Funciones de Verosimilitud , Masculino , Análisis de Secuencia de ADNRESUMEN
Genetic variations among humans occur both within and among populations and range from single nucleotide changes to multiple-nucleotide variants. These multiple-nucleotide variants are useful for studying the relationships among individuals or various population groups. The study of human genetic variations can help scientists understand how different population groups are biologically related to one another. Sequence analysis of hypervariable regions of human mitochondrial DNA (mtDNA) has been successfully used for the genetic characterization of different population groups for forensic purposes. It is well established that different ethnic or population groups differ significantly in their mtDNA distributions. In the last decade, very little research has been conducted on mtDNA variations in the Indian population, although such data would be useful for elucidating the history of human population expansion across the world. Moreover, forensic studies on mtDNA variations in the Indian subcontinent are also scarce, particularly in the northern part of India. In this report, variations in the hypervariable regions of mtDNA were analyzed in the Yadav population of Haryana. Different molecular diversity indices were computed. Further, the obtained haplotypes were classified into different haplogroups and the phylogenetic relationship between different haplogroups was inferred.
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Human mitochondrial DNA (mtDNA) is routinely analysed for pathogenic mutations, evolutionary studies, estimation of time of divergence within or between species, phylogenetic studies and identification of degraded remains. The data on various regions of human mtDNA has added enormously to the knowledge pool of population genetics as well as forensic genetics. The displacement-loop (D-loop) in the control region of mtDNA is rated as the most rapidly evolving part, due to the presence of variations in this region. The control region consists of three hypervariable regions. These hypervariable regions (HVI, HVII and HVIII) tend to mutate 5-10 times faster than nuclear DNA. The high mutation rate of these hypervariable regions is used in population genetic studies and human identity testing. In the present data, potentially informative hypervariable regions of mitochondrial DNA (mtDNA) i.e. HVI (np 16024-16365), HVII (np 73-340) and HVIII (np 438-576) were estimated to understand the genetic diversity amongst Brahmin population of Haryana. Blood samples had been collected from maternally unrelated individuals from the different districts of Haryana. An array of parameters comprising of polymorphic sites, transitions, transversions, deletions, gene diversity, nucleotide diversity, pairwise differences, Tajima's D test, Fu's Fs test, mismatch observed variance and expected heterozygosity were estimated. The observed polymorphisms with their respective haplogroups in comparison to rCRS were assigned.
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Most often the newspapers and bulletin come out with voluminous cases of deaths due to drowning. At the same time an ample section of such cases encompass entomological evidences, that can be scrutinized as a very useful parameter in estimating post mortem interval (PMI). This research database is an outcome of a 2 years reflective study, based on an assessment of records related to human deaths due to drowning. The drowned bodies from various districts of Haryana are sent to PGIMS (Post Graduate Institute of Medical Sciences), Rohtak. The study took the data of year 2015-2016 into consideration. All the cases were reviewed and summarized in terms of monthly occurrence of total cases, age differentiation, gender differentiation and month wise occurrence of entomological evidences on the dead bodies through detailed study of post mortem findings. This data will lead to an insight into the magnitude of drowning deaths in Haryana along with the usage of entomological data for determining Post Mortem Submersion Intervals (PMSI).
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With its high seed oil content, the mustard family plant Camelina sativa has gained attention as a potential biofuel source. As a bioenergy crop, camelina has many advantages. It grows on marginal land with low demand for water and fertilizer, has a relatively short life cycle, and is stress tolerant. As most other crop seed oils, camelina seed triacylglycerols (TAGs) consist of mostly long, unsaturated fatty acyl moieties, which is not desirable for biofuel processing. In our efforts to produce shorter, saturated chain fatty acyl moieties in camelina seed oil for conversion to jet fuel, a 12:0-acyl-carrier thioesterase gene, UcFATB1, from California bay (Umbellularia californica Nutt.) was expressed in camelina seeds. Up to 40% of short chain laurate (C12:0) and myristate (C14:0) were present in TAGs of the seed oil of the transgenics. The total oil content and germination rate of the transgenic seeds were not affected. Analysis of positions of these two fatty acyl moieties in TAGs indicated that they were present at the sn-1 and sn-3 positions, but not sn-2, on the TAGs. Suppression of the camelina KASII genes by RNAi constructs led to higher accumulation of palmitate (C16:0), from 7.5% up to 28.5%, and further reduction of longer, unsaturated fatty acids in seed TAGs. Co-transformation of camelina with both constructs resulted in enhanced accumulation of all three medium-chain, saturated fatty acids in camelina seed oils. Our results show that a California bay gene can be successfully used to modify the oil composition in camelina seed and present a new biological alternative for jet fuel production.
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Brassicaceae/genética , Brassicaceae/metabolismo , Aceites de Plantas/metabolismo , Semillas/metabolismo , Triglicéridos/química , Triglicéridos/metabolismo , 3-Oxoacil-(Proteína Transportadora de Acil) Sintasa/deficiencia , 3-Oxoacil-(Proteína Transportadora de Acil) Sintasa/genética , Plantas Modificadas Genéticamente , Interferencia de ARN , Tioléster Hidrolasas/genética , Umbellularia/enzimología , Umbellularia/genéticaRESUMEN
With the advances in the field of synthetic biology, there is an increasing demand for multi-gene cloning technologies. Molecular cloning to generate multi-gene constructs can be performed by restriction digestion, or by recombination-based cloning strategies such as Gateway(®). This chapter details cloning, transformation, and selection procedures involved in generation of multi-gene expressing transgenic plants. Methods are described for cloning five distinct promoter-reporter fusion constructs into the PC-GW-BAR vector (from the PC-GW vector series) using Gateway(®) technology and meganuclease sites. Further, transformation and selection methods are described for the biofuel crop Camelina sativa from the Brassicaceae family. These methods would be constructive toward generating multi-gene expressing plants for simultaneous expression analysis of five promoters in a short time period.
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Clonación Molecular/métodos , Plantas Modificadas Genéticamente/genética , Regiones Promotoras Genéticas/genética , Regulación de la Expresión Génica de las Plantas/genética , Vectores GenéticosRESUMEN
The gravitropic bending in plant roots is caused by asymmetric cell elongation. This requires an asymmetric increase in cell surface and therefore plasma membrane components such as lipids, sterols, and membrane proteins. We have identified an early gravity-regulated protein in Arabidopsis thaliana root apices that binds stigmasterol and phosphoethanolamines. This root-specific protein interacts with the membrane transport protein synaptotagmin-1 and was therefore named InteractoR Of SYnaptotagmin1 (ROSY1). While interactions between ML-domain proteins with membrane transport proteins and their impact have been reported from animal cell systems, this is the first report of such an interaction in a plant system. Homozygous mutants of ROSY1 exhibit decreased basipetal auxin transport, a faster root gravitropic response, and an increase in salt stress tolerance. Our results suggest that ROSY1 plays a role in root gravitropism, possibly by facilitating membrane trafficking and asymmetric cell elongation via its interaction with synaptotagmin-1.