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Embryonic stem cells (ESCs) have proven to be a great in vitro model that faithfully recapitulates the events that occur during in vivo embryogenesis, making them a unique tool to study the cellular and molecular mechanisms that define tissue specification during embryonic development. Livestock ESCs are particularly attractive and have broad prospects including drug selection and human disease modeling, improvement of reproductive biotechniques and agriculture-related applications such as production of genetically modified animals. While mice and human ESCs have been established many years ago, no significant advances were made in livestock species until recently. Nowadays, livestock ESCs are available from cattle, pigs, sheep, horses and rabbits with different states of pluripotency. In this review, we summarize the current advances on livestock ESCs establishment and maintenance along with their present and future applications.(AU)

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We studied the complete chloroplast genome of Gomphocarpus siniacus and Duvalia velutina from Asclepiadoideae subfamily; due to their medicinal importance and distribution worldwide their interest became high. In this study we analyzed the complete chloroplast genomes of G. siniacus and D. velutina using Illumina sequencing technology. The sequences were compared with the other species from Apocynaceae family. The complete genome of G. siniacus is 162,570 bp while D. velutina has154, 478 bp in length. Both genomes consist of 119 genes; encode 31 tRNA genes, and eight rRNA genes. Comparative studies of the two genomes showed variations in SSR markers in which G. siniacus possesses 223 while D. velutina has 186. This could be used for barcoding in order to aid in easy identification of the species. Phylogenetic analysis on the other hand reaffirms the tribal position of G. siniacus in Asclepiadeae and D. velutina in Ceropegieae. These findings could be used in subsequent research studies of angiosperms identification, genetic engineering, herb genomics and phylogenomic studies of Apocynaceae family.

Estudamos o genoma completo do cloroplasto de Gomphocarpus siniacus e Duvalia velutina da subfamília Asclepiadoideae. Em razão de sua importância medicinal e distribuição em todo o mundo, o seu interesse tornou-se elevado. Neste estudo, analisamos os genomas completos de cloroplastos de G. siniacus e D. velutina usando a tecnologia de sequenciamento Illumina. As sequências foram comparadas com as demais espécies da família Apocynaceae. O genoma completo de G. siniacus tem 162.570 pb, enquanto D. velutina tem 154.478 pb de comprimento. Ambos os genomas consistem em 119 genes e codificam 31 genes de tRNA e 8 genes de rRNA. Estudos comparativos dos dois genomas mostraram variações nos marcadores SSR em que G. siniacus possui 223, enquanto D. velutina possui 186. Isso poderia ser usado para código de barras para facilitar a identificação das espécies. A análise filogenética, por outro lado, reafirma a posição tribal de G. siniacus em Asclepiadeae e D. velutina em Ceropegieae. Esses achados poderão ser utilizados em pesquisas posteriores de identificação de angiospermas, engenharia genética, genômica de ervas e estudos filogenômicos da família Apocynaceae.

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The number of food products with genetically modified (GM) crops on the global market has increased due to advancements in genetic engineering technology. Legislation regulating the labeling and use of GM crops has increased considerably worldwide to provide consumers with health and safety assurance. It is still unclear whether genetically modified organisms (GMOs) are present in the food market of the Kingdom of Saudi Arabia due to a lack of scientific studies. This work was planned to detect GM rice and GM potatoes in the Saudi food market. One hundred non-labeled rice and rice product samples and 50 potato and potato samples were collected randomly from different market sites of Makkah, Riyadh and Jeddah during 2022-2023. The cetyl trimethyl ammonium bromide (CTAB) method was used to extract DNA. Viviants DNA extraction kit was used to extract DNA from rice starch and potato chips. To find GMOs in samples, CMOScreen 35S and NOS test kits were utilized. DNA-based qualitative and quantitative approaches were used to screen targets for PCR detection of GM rice sequences. The results indicated that 32 (32%) rice samples were positive for CaMV 35S promoter, while no positive result was detected for the NOS terminator. Besides, 30% of potato samples were positive for the CaMV 35S promoter, and the same samples were positive for the presence of the Cry V gene. It could be concluded that there were GM rice and potatoes in the Kingdom of Saudi Arabia's food markets. Establishing strong regulations and certified laboratories to monitor genetically modified foods (GMF) or crops in the Saudi market is recommended.

O número de produtos alimentícios com culturas geneticamente modificadas (GM) no mercado global cresceu devido aos avanços na tecnologia de engenharia genética. A legislação que regulamenta a rotulagem e o uso de culturas GM aumentou consideravelmente em todo o mundo para fornecer aos consumidores garantia de saúde e segurança. Ainda não está claro se organismos geneticamente modificados (OGM) estão presentes no mercado de alimentos do Reino da Arábia Saudita em razão da falta de estudos científicos. Este trabalho foi planejado para detectar arroz e batatas GM no mercado de alimentos saudita. Cem amostras de arroz e produtos de arroz não rotulados e 50 amostras de batata e produtos à base de batata foram coletadas aleatoriamente de diferentes mercados de Meca, Riad e Jeddah durante 2022-2023. O método de brometo de cetiltrimetilamônio (CTAB) foi usado para extrair DNA. O kit de extração de DNA Viviants foi usado para extrair DNA de amido de arroz e batatas fritas. Para encontrar OGMs em amostras, foram utilizados os kits de teste CMOScreen 35S e NOS. Abordagens qualitativas e quantitativas baseadas em DNA foram usadas para rastrear alvos para detecção de PCR de sequências de arroz GM. Os resultados indicaram que 32 (32%) amostras de arroz foram positivas para o promotor CaMV 35S, enquanto nenhum resultado positivo foi detectado para o terminador NOS. Além disso, 30% das amostras de batata foram positivas para o promotor CaMV 35S, e as mesmas amostras foram positivas para a presença do gene Cry V. Pode-se concluir que havia arroz e batatas GM nos mercados de alimentos do Reino da Arábia Saudita. É recomendável estabelecer regulamentações rigorosas e laboratórios certificados para monitorar alimentos geneticamente modificados (GMF) ou safras no mercado saudita.

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One of the most serious problems worldwide is heavy metal (HM) pollution. HMs can have a toxic effect on human health and thus cause serious diseases. To date, several methods have been used to clean environments contaminated by HMs, but most of them are expensive, and it is difficult to achieve the desired result. Phytoremediation is currently an effective and affordable processing solution used to clean and remove HMs from the environment. This review article discusses in detail the technology of phytoremediation and mechanisms of HM absorption. In addition, methods are described using genetic engineering of various plants to enhance the resistance and accumulation of HMs. Thus, phytoremediation technology can become an additional aid to traditional methods of purification.

Um dos problemas mais graves em todo o mundo é a poluição por metais pesados (HMs). Os HMs podem ter um efeito tóxico na saúde humana e, assim, causar doenças graves. Até o momento, vários métodos têm sido utilizados para limpar ambientes contaminados por HMs, mas a maioria deles é cara, sendo difícil alcançar o resultado desejado. A fitorremediação é, atualmente, uma solução de processamento eficaz e acessível usada para limpar e remover HMs do ambiente. Este artigo de revisão discute em detalhes a tecnologia de fitorremediação e os mecanismos de absorção de HMs. Além disso, são descritos métodos que utilizam a engenharia genética de várias plantas para aumentar a resistência e o acúmulo de HMs. Assim, a tecnologia de fitorremediação pode se tornar uma ajuda adicional aos métodos tradicionais de purificação.

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The objective of this review is to bring information about innovations and technologies that, through genetic improvement, are being used to improve the sustainability and productivity of agricultural crops, improve human nutrition, as well as conservation and decontamination of soils. Bioremediation consists of using microorganisms that have the ability to modify or decompose certain pollutants, with the possibility of increasing their activity through genetic engineering, building new strains for the transformation of pollutants into inert substances. Genetic improvement is seeking to develop cultivars that are more tolerant to periods of water deficit. Plant biofortification consists of varieties of improved plants that have a higher content of vitamins and minerals, which are obtained through genetic improvement. Thus, biotechnology is once again essential for world agricultural production and can bring a series of other benefits to society.(AU)

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This article provides an overview of assisted reproductive technologies (ART) and genome engineering to improve livestock production systems for the contribution of global sustainability. Most ruminant production systems are conducted on grassland conditions, as is the case of South American countries that are leaders in meat and milk production worldwide with a well-established grass-feed livestock. These systems have many strengths from an environmental perspective and consumer preferences but requires certain improvements to enhance resource efficiency. Reproductive performance is one of the main challenges particularly in cow-calf operations that usually are conducted under adverse conditions and thus ART can make a great contribution. Fixed-time artificial insemination is applied in South America in large scale programs as 20 to 30% of cows receive this technology every year in each country, with greater calving rate and significant herd genetic gain occurred in this region. Sexed semen has also been increasingly implemented, enhancing resource efficiency by a) obtaining desired female replacement and improving animal welfare by avoiding newborn male sacrifice in dairy industry, or b) alternatively producing male calves for beef industry. In vitro embryo production has been massively applied, with this region showing the greatest number of embryos produced worldwide leading to significant improvement in herd genetics and productivity. Although the contribution of these technologies is considerable, further improvements will be required for a significant livestock transformation and novel biotechnologies such as genome editing are already available. Through the CRISPR/Cas-based system it is possible to enhance food yield and quality, avoid animal welfare concerns, overcome animal health threats, and control pests and invasive species harming food production. In summary, a significant enhancement in livestock productivity and resource efficiency can be made through reproductive technologies and genome editing, improving at the same time profitability for farmers, and global food security and sustainability.(AU)

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Currently, gonadotropin products (follicle stimulating hormone, FSH, and luteinizing hormone, LH) used in animal reproduction are produced by extraction and purification from abattoir-derived pituitary glands. This method, relying on animal-derived materials, carries the potential risk of hormone contamination and pathogen transmission. Additionally, chorionic gonadotropins are extracted from the blood of pregnant mares (equine chorionic gonadotropin; eCG) or the urine of pregnant women (human chorionic gonadotropin; hCG). However, recent advancements have introduced recombinant gonadotropins for assisted animal reproduction therapies. The traditional use of FSH for superovulation has limitations, including labor requirements and variability in superovulation response, affecting the success of in vivo (SOV) and in vitro (OPU/IVEP) embryo production. FSH treatment for superstimulation before OPU can promote the growth of a homogenous follicular population and the recovery of competent oocytes suitable for IVEP procedures. At present, a single injection of a preparation of long-acting bovine recombinant FSH (rFSH) produced similar superovulation responses resulting in the production of good-quality in vivo and in vitro embryos. Furthermore, the treatment with eCG at FTAI protocol has demonstrated its efficacy in promoting follicular growth, ovulation, and P/AI, mainly in heifers and anestrous cows. Currently, treatment with recombinant glycoproteins with eCG-like activity (r-eCG) have shown promising results in increasing follicular growth, ovulation, and P/AI in cows submitted to P4/E2 -based protocols. Bovine somatotropin (bST) is a naturally occurring hormone found in cows. Recombinant bovine somatotropin (rbST), produced through genetic engineering techniques, has shown potential in enhancing reproductive outcomes in ruminants. Treatment with rbST has been found to improve P/IA, increase donor embryo production, and enhance P/ET in recipients. The use of recombinant hormones allows to produce non-animal-derived products, offering several advantages in assisted reproductive technologies for ruminants. This advancement opens up new possibilities for improving reproductive efficiency and success rates in the field of animal reproduction.(AU)

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This study aimed to carry out the morphological characterization of the remaining specimens of the Curraleiro horse in municipalities of the state of Goiás, Brazil. Forty male horses were evaluated using the age of five years as a criterion. Sixteen linear measurements and 13 zootechnical indices were obtained. Subsequently, the mean, standard deviation, minimum, maximum, and covariances of these measures and indices were obtained using the software IBM SPSS Statistics for Windows. The measures withers height (WH), croup height (CH), midback height (MH), sternum-to-ground height (SH), chest index (CI), and estimated weight (W) allowed characterizing Curraleiro horses as small-sized, light, and fast, with proportional measures. The dactyl-thoracic index (DTI), body index (BI), conformation index (CFI), load index 1 and 2 (LOI1 and LOI2), and compactness index 1 and 2 (COI1 and COI2) showed that the Curraleiro horse has an intermediate capacity for speed and strength, bearing considerable weight on the back, with saddling aptitude and fast work. These results represent the beginning of the formation of a database that may contribute to future studies and the conservation of the Curraleiro horse in the state of Goiás.(AU)

O objetivo deste trabalho foi a realização da caracterização morfológica de espécimes remanescentes do cavalo Curraleiro em municípios do estado de Goiás, Brasil. Foram avaliados 40 cavalos machos, tendo como critério a idade a partir de cinco anos, dos quais se obteve 16 medidas lineares e 13 índices zootécnicos. Posteriormente, a média, o desvio padrão, o mínimo, o máximo e as covariâncias dessas medidas e índices foram obtidos por meio da utilização do software IBM SPSS Statistics for Windows. As medidas altura da cernelha (AC), altura da garupa (AG), altura do costado (ACost), vazio subesternal (VAZ), índice peitoral (IP) e peso estimado (P) permitiram caracterizar os cavalos Curraleiros como um grupo de pequeno porte, leve, veloz e com medidas proporcionais. Os índices dáctilo-torácico (IDT), corporal (IC), de conformação (ICF), de carga 1 e 2 (ICG1 e ICG2) e de compacidade 1 e 2 (ICO1 e ICO2) demonstraram que o cavalo Curraleiro tem capacidade intermediária para velocidade e força, suportam considerável peso sobre o dorso, e possuem aptidão para sela e tração ligeira. Tais resultados representam o início da formação de uma base de dados que pode contribuir para estudos futuros e a conservação do cavalo Curraleiro no estado de Goiás.(AU)

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Biochemical and physiological parameters, growth, and yield of field crops especially salt sensitive crops like chickpea are affected adversely by salinity in arid to semi-arid regions. To investigate the effect of different salinity levels on growth, biochemical and physiological parameters of chickpea genotypes, a pot experiment following CRD, two factor factorial design, was conducted in the glasshouse at the Institute of Biotechnology and Genetic Engineering, The University of Agriculture, Peshawar, Pakistan. Ten (10) kg of soil was filled in each pot and salinity levels were maintained @ S0= 0 mM NaCl, S1= 50 mM NaCl, S2= 100 mM NaCl and S3= 150 mM by applying NaCl and 5 genotypes of chickpea (KK-2, Bhakkar-2011, Bittle-98, Punjab-2008, and CM-98) were used. At crop maturity, growth parameters, physiological, biochemical, and ionic parameters were measured using standard analysis procedures. Salinity reduced the growth and yield of all genotypes, but the rate of decrease was different among the genotypes tested. From the results, a decrease in K concentration, K/Na ratio, transpiration rate, stomatal conductance, N, and P was observed in all genotypes with the increase in salinity. An increase in salinity level increased the proline content (35.45%), crude protein (42%), H2O2 (19%), lipid peroxidation (62%), carbohydrates (23.22%), and Na+ concentration (137%). The highest level of salinity, 150 mM NaCl has exhibited the highest salinity stress in all parameters. Genotype KK-2 and Bhakkar-11 showed a lower rate of relative decrease in yield (4.5 and 12%), K+/Na+ ratio (23.34 and 11.47%), and K+ concentration (7.9 and 11%), respectively, and the lowest relative increase in Na+ accumulation (20.3 and 0.48%), @ 50 mM salinity compared to control. Genotype KK-2 and Bhakkar-11 proved better @ 50mM salinity. The findings suggest that the critical level of the salinity must be kept in mind and the salt-tolerant genotypes should be cultivated in salt affected soils.

Parâmetros bioquímicos e fisiológicos, crescimento e rendimento de culturas de campo, especialmente culturas sensíveis ao sal, como grão-de-bico, são afetados negativamente pela salinidade em regiões áridas e semiáridas. Para investigar o efeito de diferentes níveis de salinidade no crescimento, parâmetros bioquímicos e fisiológicos de genótipos de grão-de-bico, um experimento em pote seguindo CRD, delineamento fatorial de dois fatores, foi conduzido na estufa do Instituto de Biotecnologia e Engenharia Genética, Universidade de Agricultura, Peshawar, Paquistão. Dez kg de solo foram preenchidos em cada vaso e os níveis de salinidade foram mantidos @ S0 = 0 mM NaCl, S1 = 50 mM NaCl, S2 = 100 mM NaCl e S3 = 150 mM aplicando NaCl e 5 genótipos de grão-de-bico (KK-2, Bhakkar-2011, Bittle-98, Punjab-2008 e CM-98). Na maturidade da cultura, parâmetros de crescimento, parâmetros fisiológicos, bioquímicos e iônicos foram medidos usando procedimentos de análise padrão. A salinidade reduziu o crescimento e a produtividade de todos os genótipos, mas a taxa de decréscimo foi diferente entre os genótipos testados. A partir dos resultados, observou-se diminuição da concentração de K, razão K/Na, taxa de transpiração, condutância estomática, N e P em todos os genótipos com o aumento da salinidade. Um aumento no nível de salinidade aumentou o teor de prolina (35,45%), proteína bruta (42%), H2O2 (19%), peroxidação lipídica (62%), carboi- dratos (23,22%) e concentração de Na+ (137%). O nível mais alto de salinidade, 150 mM NaCl, exibiu o maior estresse de salinidade em todos os parâmetros. Os genótipos KK-2 e Bhakkar-11 apresentaram menor taxa de diminuição relativa no rendimento (4,5 e 12%), razão K+/Na+ (23,34 e 11,47%) e concentração de K+ (7,9 e 11%), respectivamente, e menor aumento relativo no acúmulo de Na+ (20,3 e 0,48%), @ 50 mM de salinidade comparado ao controle. Os genótipos KK-2 e Bhakkar-11 se mostraram melhores @ 50mM de salinidade. Os resultados sugerem que o nível crítico de salinidade deve ser mantido em mente e os genótipos tolerantes ao sal devem ser cultivados em solos afetados pelo sal.

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O desenvolvimento de estudos genéticos e de microdispositivos biológicos tem proporcionado a ampliação do conhecimento sobre os complexos eventos que envolvem a reprodução animal. O desafio ainda é imensurável, mas a criação e surgimentos de novas perspectivas para a pesquisa básica tem-se feito presente. Neste trabalho revisamos de maneira suscinta algumas abordagens recentes, utilizadas pela pesquisa básica, sobretudo com o objetivo de lançar luz sobre o desenvolvimento folicular e oocitário. Dessa forma, essa revisão pretende fornecer uma visão geral do uso das tecnologias ômicas e sistema de microfluídica como auxiliadores na compreensão da foliculogênese. Adicionalmente serão apresentadas particularidades inerentes à fisiologia da gametogênese, que incluem ação de microorganismos e mitocôndrias, além do importante papel da comunicação intercelular através das vesículas extracelulares.

The development of genetic studies and biological microdevices has expanded knowledge about the complex events involving animal reproduction. The challenge is still immeasurable, but the creation and emergence of new perspectives for basic research have been present. This paper briefly reviews some recent approaches used in basic research, mainly to shed light on follicular and oocyte development. Thus, this review intends to provide an overview of the use of omics technologies and microfluidics systems as aids in understanding folliculogenesis. Also, it will present particulars inherent in the physiology of gametogenesis, which include microorganisms and mitochondria, in addition to the important role of intercellular communication through extracellular vesicles.

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O transplante de espermatogônias tronco (SSCs, do inglês Spermatogonial Stem Cell) é uma biotecnologia que consiste na transferência de células tronco testiculares de um doador fértil para um receptor cuja espermatogênese endógena foi suprimida. Essa técnica pode ser aplicada para a produção de machos que gerem uma progênie com características genotípicas do doador selecionado. Especialmente na bovinocultura, tanto de leite como de corte, o transplante de SSCs tem o potencial de substituir a inseminação artificial (IA). Pode-se também, colocar SSCs de um mesmo doador (de genética superior) em mais de um receptor o que aumentaria o número de filhos desse doador. Além disso, possui outras aplicações como a restauração da fertilidade em homens após o tratamento de câncer, conservação de espécies ameaçadas de extinção e tratamento de causas específicas de infertilidade. Assim, com esta revisão tem-se como propósito discorrer acerca de uma biotecnologia da reprodução que permitirá a propagação do valor genético de doadores de sêmen considerados de alto valor zootécnico.

Spermatogonial Stem Cell (SSCs) transplantation is a biotechnology that consists in the transfer of testicular stem cells from a fertile donor to a recipient whose endogenous spermatogenesis has been depleted. This technique can be applied to the production of males that generate a progeny with genotypic characteristics of the selected donor. Especially in beef and dairy cattle, SSCs transplantation has the potential to replace artificial insemination (AI). In addition, it has other applications such as restoring fertility in human species after cancer treatment, conserving endangered species and treating specific causes of infertility. Thus, this aim of this review is to discuss the perspectives of reproductive biotechnology that allows the propagation of the genetic material of high pedigree males.

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This study evaluated the physicochemical and morphological properties of a marine sponge protein extract (PE) using scanning electron microscopy (SEM), Energy dispersive X-ray spectroscopy (EDS), analysis of mass loss and pH and in vitro and in vivo. Scanning electron microscopy showed that PE fibers present a granular aspect and irregular structure and the element carbon followed by oxygen was detected in the EDS analysis. Moreover, a 29% of mass loss was observed after 14 days and the pH slightly modified after 14 days. Cell viability of fibroblast cells (L929) of control and PE at a concentration of 25% demonstrated higher values compared to the groups. Osteoblast cell viability of PE at 25 and 50% was significantly higher. Comet assay on day 1 showed higher values for PE at 25%. In addition, in vivo experiments demonstrated that in the treated animals, the bone defects were filled with biomaterial particles, granulation tissue and some areas of newly formed bone. Furthermore, similar immunoexpression of Runx-2 and Cox-2 was observed. Taken together, all results suggest that PE is biocompatible, present non-citotoxicity in the in vitro studies (at the lower concentration) and in the in vivo studies and it can be considered as an alternative source of collagen for tissue engineering proposals.

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Soybean loss due to pests and pathogens is a serious problem worldwide. Soybean producers have few options to manage diseases caused by general pathogens where major genes for full resistance have not been discovered. The innate defense of soybean plants could be enhanced by improving content and composition of lignin by genetic engineering of the phenylpropanoid pathway.We used a novel technique of germ-line genetic transformation of soybean plants via natural pollen tubes as vectors. This technique uses Agrobacterium tumefaciensto mediate transfer of genes of interest to the zygote to introduce the key lignification genes (PtMYB4, PAL5, F5H, CAD1) into soybean genome. We observed 5.6% average transformation efficiency in the first generation of transgenic plants and in the second generation the presence of the transgene constructs was confirmed in more than 50% (for CsVMV/PtMYB4sens, 35SVTM/PAL5, C4H/F5H, CsVMV/CAD1constructs) transgenic soybean lines. We confirmed the expression of the introduced genes at transcriptional level using RT-PCR and Northern blot. Functional analysis using lignin content determination and the activity of PAL5 and CAD1 enzymes demonstrated that the transgenes perform their function in planta. The proposed technique is effectiveand inexpensive and can be used to create novel stress and disease resistant soybean genotypes.

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ABSTRACT: Gene stacking refers to the introduction of two or more transgenes of agronomic interest in the same plant. The main methods for genetically engineering plants with gene stacking involve (i) the simultaneous introduction, by the co-transformation process, and (ii) the sequential introduction of genes using the re-transformation processes or the sexual crossing between separate transgenic events. In general, the choice of the best method varies according to the species of interest and the availability of genetic constructions and preexisting transgenic events. We also present here the use of minichromosome technology as a potential future gene stacking technology. The purpose of this review was to discuss aspects related to the methodology for gene stacking and trait stacking (a gene stacking strategy to combine characteristics of agronomical importance) by genetic engineering. In addition, we presented a list of crops and genes approved commercially that have been used in stacking strategies for combined characteristics and a discussion about the regulatory standards. An increased number of approved and released gene stacking events reached the market in the last decade. Initially, the most common combined characteristics were herbicide tolerance and insect resistance in soybean and maize. Recently, commercially available varieties were released combining these traits with drought tolerance in these commodities. New traits combinations are reaching the farmer's fields, including higher quality, disease resistant and nutritional value improved. In other words, gene stacking is growing as a strategy to contribute to food safety and sustainability.

RESUMO: O empilhamento gênico se refere a introdução de dois ou mais transgenes de interesse agronômico na mesma planta. Os principais métodos de produção de plantas geneticamente modificadas com empilhamento gênico envolvem (i) a introdução simultânea, pelo processo de co-transformação, e (ii) a introdução sequencial de genes, pelos processos de re-transformação ou por cruzamento entre eventos transgênicos. Em geral, a escolha do melhor método varia de acordo com a espécie de interesse e a disponibilidade de construções genéticas e eventos transgênicos preexistentes. Também é apresentado aqui o uso da tecnologia de minicromossomos como tecnologia potencial de empilhamento gênico. O objetivo desta revisão é discutir aspectos relacionados à metodologia para o empilhamento de genes a combinação de características (obtida via empilhamento de genes de interesse agronômico) via engenharia genética. Além de discutir, é apresentado uma lista de culturas e genes aprovados comercialmente que tem sido usado em estratégias de empilhamento e uma discussão sobre normas regulatórias. Um número maior de eventos com empilhamento de genes foi aprovado e liberado no mercado na última década. Inicialmente, a combinação das características de tolerância a herbicidas e resistência a insetos era a mais popular, principalmente em soja e milho. Recentemente, estas características combinadas com tolerância a seca nessas culturas foram liberadas comercialmente. Novas características combinadas estão entrando na lavoura, incluindo aumento da qualidade, resistência a doenças e aumento do valor nutricional. Em outras palavras, o empilhamento gênico está crescendo como tecnologia para contribuir para a segurança alimentar e sustentabilidade.

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With a focus on productivity, Brazil has advanced research and application of reproductive biotechnologies. However, much remains to be explored, such as in vitro embryo production and transfer in miniature breeds. Thus, the objective of the present work is to report the in vitro production of miniature bovine embryos and transfer to conventional sized cows located in Palmas/To. Nine miniature Jersey-Holstein cross breed donors were aspirated to obtain approximately 13 viable oocytes per female. The oocytes were matured, fertilized and cultured the in vitro, where the production of viable embryos was 23.2%. On day 7 of development (D7) the embryos were transferred to 17 crossbred dairy recipients and after 30 days of innovulation by ultrasound examination a conception rate of 76.4% was found. Thus, the technique of in vitro production and embryo transfer is viable for the reproduction of mini dairy cows, despite the lack of data regarding the work performed on small animals that serve as comparative parameters for the results achieved in the study.

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ABSTRACT: This paper reflects on the controversy between agroecology and modern conventional agriculture. As a counter-paradigm, agroecology criticizes and resists advances in agricultural sciences, including biotechnology. But does not offer concrete answers to the challenge of increasing food production in the world in a sustentably way. It is a view that denies the importance of modern agriculture to solve the problems of scarcity and food security and does not recognize the potential of genetic modification to decisively to promote sustainable agriculture, accelerating the overcoming of the so-called "chemical reductionist paradigm". The principal argument is that agroecology, despite being an area of ​​knowledge that can contribute to recovering natural resources and helping nature's resilience, is not an alternative to modern conventional agriculture and has been trying to prevent advances in genetic engineering, a fundamental area for sustainability and for the low carbon agriculture. In the end, based on the importance of the controversy in the History of Science, presents the reasons why agroecology does not fulfill the requirements to be an agricultural science.

RESUMO: Este artigo reflete sobre a controvérsia entre a agroecologia e a agricultura convencional moderna. Como um contra-paradigma, a agroecologia critica e resiste aos avanços nas ciências agrícolas, incluindo a biotecnologia. Mas não oferece respostas concretas ao desafio de aumentar a produção de alimentos no mundo de maneira sustentável. É uma visão que nega a importância da agricultura moderna para resolver os problemas de escassez e segurança alimentar e não reconhece o potencial da modificação genética para promover decisivamente a agricultura sustentável, acelerando a superação do chamado "paradigma reducionista químico". O principal argumento é que a agroecologia, apesar de ser uma área do conhecimento que pode contribuir para recuperar os recursos naturais e ajudar a resiliência da natureza, não é uma alternativa à agricultura convencional moderna e vem tentando impedir avanços na engenharia genética, uma área fundamental para a sustentabilidade e para a agricultura de baixo carbono. Por fim, com base na importância da controvérsia na História da Ciência, explora as razões pelas quais a agroecologia não preenche os requisitos para ser uma ciência agrária.

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Abstract The global prevalence of diabetes mellitus and other metabolic diseases is rapidly increasing. Animal models play pivotal roles in unravelling disease mechanisms and developing and testing therapeutic strategies. Rodents are the most widely used animal models but may have limitations in their resemblance to human disease mechanisms and phenotypes. Findings in rodent models are consequently often difficult to extrapolate to human clinical trials. To overcome this translational gap, we and other groups are developing porcine disease models. Pigs share many anatomical and physiological traits with humans and thus hold great promise as translational animal models. Importantly, the toolbox for genetic engineering of pigs is rapidly expanding. Human disease mechanisms and targets can therefore be reproduced in pigs on a molecular level, resulting in precise and predictive porcine (PPP) models. In this short review, we summarize our work on the development of genetically (pre)diabetic pig models and how they have been used to study disease mechanisms and test therapeutic strategies. This includes the generation of reporter pigs for studying beta-cell maturation and physiology. Furthermore, genetically engineered pigs are promising donors of pancreatic islets for xenotransplantation. In summary, genetically tailored pig models have become an important link in the chain of translational diabetes and metabolic research.

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Cell fate specification, gene expression and spatial restriction are process finely tuned by epigenetic regulatory mechanisms. At the same time, mechanical forces have been shown to be crucial to drive cell plasticity and boost differentiation. Indeed, several studies have demonstrated that transitions along different specification states are strongly influenced by 3D rearrangement and mechanical properties of the surrounding microenvironment, that can modulate both cell potency and differentiation, through the activation of specific mechanosensing-related pathways. An overview of small molecule ability to modulate cell plasticity and define cell fate is here presented and results, showing the possibility to erase the epigenetic signature of adult dermal fibroblasts and convert them into insulin-producing cells (EpiCC) are described. The beneficial effects exerted on such processes, when cells are homed on an adequate substrate, that shows “in vivo” tissue-like stiffness are also discussed and the contribution of the Hippo signalling mechanotransduction pathway as one of the mechanisms involved is examined. In addition, results obtained using a genetically modified fibroblast cell line, expressing the enhanced green fluorescent protein (eGFP) under the control of the porcine insulin gene (INS) promoter (INS-eGFP transgenic pigs), are reported. This model offers the advantage to monitor the progression of cell conversion in real time mode. All these observations have a main role in order to allow a swift scale-up culture procedure, essential for cell therapy and tissue engineering applied to human regenerative medicine, and fundamental to ensure an efficient translation process from the results obtained at the laboratory bench to the patient bedside. Moreover, the creation of reliable in vitro model represents a key point to ensure the development of more physiological models that, in turn, may reduce the number of animals used, implementing non-invasive investigations and animal welfare and protection.

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Theileria parva is the causative agent of East Coast Fever (ECF), a tick borne disease, which results in major economic losses in cattle. Major problems in dealing with this illness are the high cost of drugs, development of resistance, and absence of effective vaccines. Thus, exploiting new targets for cost effective and higher therapeutic value drugs are imperative. Glycolysis is the main pathway for generation of ATP in T. parva, given its development inside erythrocytes. Thus, the enzymes of this pathway may prove potential targets for designing new-generation anti-theilerials. Lactate dehydrogenase of T. parva (TpLDH) has the highest activity of all glycolytic enzymes and thus we selected this enzyme as the potential therapeutic target. Our study is the first to report the isolation, removal of introns through directed mutagenesis, and cloning of TpLDH and showing that amino acid insertions or deletions most notably corresponded to a 5-amino acid sequence (Asn-91A, Glu-91B, Glu-91C, Trp-91D, Asn-91E) between Ser-91 ve Arg-92 of the enzyme. This region is also present in other apicomplexan such as Babesia bovis, a pathogen of cattle and Plasmodium falciparum, a human pathogen. Providing as the attachment site for the enzyme inhibitors and not being present in LDH of respective hosts, we propose this site as an attractive drug target. The work here is expected to lead new studies on detailed structural and kinetic aspects of apicomplexan LDHs and development of new inhibitors.(AU)

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A biologia sintética é uma ciência que está crescendo em estudos de pesquisas devido à capacidade que tem de criar organismos e micro-organismos vivos que produzam uma função que naturalmente não realizam. Pesquisadores têm se mostrado eficientes na criação de genes em laboratórios por meio do código da vida e sintetizadores, padronizando genomas com um determinado fim. A agronomia, química e farmácia têm sido as grandes áreas que essa ciência vem sendo desenvolvida. Portanto, o presente estudo teve como objetivo realizar uma pesquisa bibliográfica relatando alguns desenvolvimentos produzidos pelos pesquisadores das áreas afins. Com a biologia sintética foi possível a utilização de bactérias contra células cancerígenas, bactérias que produz biocombustível, assim como antibióticos produzidos por bactérias, entre outros.(AU)

The number of research studies in synthetic biology is increasing due to its capacity of creating live organisms and micro-organisms that can produce features that are not naturally produced. Researchers have been efficient in creating genes in laboratory through the code of life and synthesizers, standardizing genomes for a given purpose. Agriculture, chemistry and pharmacy have been the main areas developed by this science. Therefore, this study had the purpose of performing a literature review reporting some of the developments produced by researchers in the correlated areas. Synthetic biology enabled the use of bacteria against cancer cells, bacteria that can produce biofuel, as well as bacteria-produced antibiotics, among others.(AU)

La biología sintética es una ciencia que está creciendo en estudios de investigación debido a la capacidad que tiene de crear organismos y microorganismos vivos que produzcan una función que naturalmente no realizan. Investigadores se han mostrado eficientes en la creación de genes en laboratorios a través del código de la vida y sintetizadores, estandarizando genomas con un determinado fin. La agronomía, química y farmacia han sido las grandes áreas que esta ciencia viene siendo desarrollada. Por lo tanto, el presente estudio tuvo como objetivo realizar una investigación bibliográfica relatando algunos desarrollos producidos por los investigadores de las áreas afines. Con la biología sintética fue posible el uso de bacterias contra células cancerígenas, bacterias que producen biocombustibles, así como antibióticos producidos por bacterias, entre otros.(AU)