Novel agents targeting leukemia cells and immune microenvironment for prevention and treatment of relapse of acute myeloid leukemia after allogeneic hematopoietic stem cell transplantation

Relapse remains the worst life-threatening complications after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in patients with acute myeloid leukemia (AML), whose prognosis has been historically dismal. Given the rapid development of genomics and immunotherapies, the interference strategies for AML recurrence have been changing these years. More and more novel targeting agents that have received the U.S. Food and Drug Administration (FDA) approval for

The mechanism of plant gall induction by insects: revealing clues, facts, and consequences in a cross-kingdom complex interaction

Galls are defined as modifications of the normal developmental design of plants, produced by a specific reaction to the presence and activity of a foreign organism. Although different organisms have the ability to induce galls in plants, insect-induced galls are the most elaborate and diverse. Some hypotheses have been proposed to explain the induction mechanism of plant galls by insects. The most general hypothesis suggests that gall formation is triggered by the action of chemical substances secreted by the gall inducer, including plant growth regulators such as auxins, cytokinins, indole-3-acetic acid (IAA), and other types of compounds. However, the mode of action of these chemical substances and the general mechanism by which the insect could control and manipulate plant development and physiology is still not known. Moreover, resulting from the complexity of the induction process and development of insect galls, the chemical hypothesis is very unlikely a complete explanation of the mechanism of induction and morphogenesis of these structures. Previous and new highlights of insect gall systems with emphasis on the induction process were analyzed on the basis of the author's integrated point of view to propose a different perspective of gall induction, which is provided in this article. Due to the extraordinary diversity of shapes, colors, and complex structures present in insect galls, they are useful models for studying how form and structure are determined at the molecular level in plant systems. Furthermore, plant galls constitute an important source of material for the study and exploration of new chemical substances of interest to humans, due to their physiological and adaptive characteristics. Considering the finely tuned control of morphogenesis, structural complexity, and biochemical regulation of plant galls induced by insects, it is proposed that an induction mechanism mediated by the insertion of exogenous genetic elements into the genome of plant gall cells could be involved in the formation of this kind of structure through an endosymbiotic bacterium.

Las agallas se definen como modificaciones del diseño y desarrollo normal de las plantas debido a una reacción específica a la presencia y actividad de un organismo foráneo. Aunque diferentes grupos de organismos tienen la habilidad de inducir agallas en plantas, las agallas inducidas por insectos son las más elaboradas y diversas. Algunas hipótesis han sido propuestas para explicar el mecanismo de inducción de las agallas de insectos. La hipótesis más general sugiere que la formación de las agallas es disparada por la acción de sustancias químicas secretadas por el insecto inductor, incluyendo reguladores de plantas como auxinas, citoquininas, ácido-3-indolacético (AIA) o bien otros tipos de compuestos. No obstante, el modo de acción de estas sustancias químicas y el mecanismo general por medio del cual el insecto podría controlar y manipular el desarrollo y fisiología de la planta es aún desconocido. Más aún, como resultado de la complejidad del proceso de inducción y desarrollo de las agallas de plantas inducidas por insectos, la hipótesis química es una explicación insuficiente e incompleta en relación con el mecanismo de inducción y morfogénesis de estas estructuras. Previas y nuevas evidencias relacionadas con el sistema de agallas de insectos, con énfasis en el proceso de inducción, fueron analizadas desde un punto de vista integral del autor para proponer en este artículo una perspectiva diferente sobre la inducción de este tipo de estructuras. Debido a la extraordinaria diversidad de formas, colores y estructuras complejas presentes en las agallas de insectos, las mismas constituyen modelos útiles para estudiar cómo la forma y la estructura son determinadas a nivel molecular en los sistemas vegetales. Además, las agallas de plantas son un importante origen de material para el estudio y exploración de nuevas sustancias químicas de interés humano, debido a las características fisiológicas y adaptativas que presentan. Considerando el control fino del proceso de morfogénesis, regulación bioquímica y complejidad estructural de las agallas de insectos, se propone que un mecanismo de inducción mediado por la inserción de elementos genéticos exógenos dentro del genoma de las células de la planta que forman la agalla podría estar involucrado en la formación de este tipo de estructuras, vía una bacteria endosimbiótica.

Real-time monitoring of translocation of selected type-III effectors from Xanthomonas oryzae pv. oryzae into rice cells

Type-III (T3) effectors PthXo1 and AvrXa10 of Xanthomonas oryzae pv. oryzae are translocated into rice cells to inducevirulence and avirulence on susceptible- and resistant-rice varieties Nipponbare and IRBB10, respectively. The translocation needs the bacterial T3 translocator Hpa1 and rice Oryza sativa plasma membrane protein OsPIP1;3. Here, weemployed the b-lactamase (BlaM) reporter system to observe PthXo1 and AvrXa10 translocation. The system wasestablished to monitor effectors of animal-pathogenic bacteria by quantifying the BlaM hydrolysis product [P] and fluorescence resonance energy transfer (FRET) of the substrate. The feasibility of the BlaM reporter in rice protoplasts wasevaluated by three criteria. The first criterion indicated differences between both [P] and FRET levels among wild types andOsPIP1;3-overexpressing and OsPIP1;3-silenced lines of both Nipponbare and IRBB10. The second criterion indicateddifferences between [P] and FRET levels in the presence and absence of Hpa1. The last criterion elucidated the coincidenceof PthXo1 translocation with induced expression of the PthXo1 target gene in protoplasts of Nipponbare and the coincidence of AvrXa10 translocation with induced expression of the AvrXa10 target gene in protoplasts of IRBB10. Theseresults provide an experimental avenue for real-time monitoring of bacterial T3 effector translocation into plant cells with apathological consequence.

Effector Pathways of Toll-like Receptor-inducible Innate Immune Responses in Macrophages

Toll-like receptors (TLR) are well-characterized pattern recognition receptors that can recognize and respond to diverse pathogen-associated or danger-associated molecular patterns during infection. TLR signaling in macrophages triggers in the intracellular signaling pathways through the recruitment of various adaptor and signaling proteins, and results in the activation of effector mechanisms and pathways that are important for host defense to intracellular bacteria. Effector mechanisms include inflammatory responses, cytokine generation, production of reactive oxygen species, and antimicrobial proteins. Accumulating studies showed that autophagy is a key pathway in the maintenance of homeostasis and housekeeping functions during infection and inflammation. In this review, we summarize the major effector pathways and mechanisms in the activation of TLR-inducible innate immune responses in macrophages. In addition, we focus the emerging evidence of crosstalk between autophagy and TLR-mediated signaling in terms of effector function of innate immune responses. A better understanding of effector functions by the activation of TLR-mediated signaling cascades contributes to the development of new therapeutics and vaccines against various intracellular pathogenic infections.

Botulinum toxin:novel strategy focusing on the effector / 中华神经科杂志

As a local and effective denervation method,botulinum toxin has been applied in more and more fields.Practitioners who use botulinum toxin should consider the general situation of patients based on the view of effectors,weigh the pros and cons and set reasonable goals before injection,referring to the Chinese expert consensus.

Research progress on the relationship between Rho GTPases and breast cancer / 天津医药

Breast cancer is one of the major causes of death in women,and its incidence has been increasing year after year.The Rho GTPases,their regulatory proteins and Rho GTPases play an important role in promoting the occurrence and distant metastasis of breast cancer.Here we summarized the current knowledge of the regulation network of Rho GTPases,their regulatory proteins and Rho GTPases on the occurrence and development of breast cancer,and targeted therapy for RHO GTP enzyme pathway in breast cancer.

Expresión de efectores RXLR en el linaje clonal EC-1 de Phytophthora infestans en Perú / Expression of RXLR effectors in the EC-1 clonal lineage of Phytophthora infestans in Peru

La papa (Solanum tuberosum L.) es en nuestros días uno de los cultivos alimenticios más importantes. Esta es propensa a la enfermedad de Tizón Tardío, causada por el oomiceto patógeno Phytophthora infestans, el cual secreta cientos de efectores que actúan como factores de virulencia. Poco se conoce sobre la diversidad de genes de virulencia de las cepas pertenecientes al linaje de reproducción clonal EC-1. En el presente estudio, mediante el secuenciamiento del transcriptoma de la interacción de la papa y P. infestans durante los primeros días después de la infección en las hojas de papa, se identificó la expresión diferencial de genes efectores tipo RXLR en dos cepas de P. infestans EC-1, siendo confirmados por qRT-PCR. Estas cepas, aisladas de papas cultivadas en el centro de los Andes peruanos, tienen diferentes patrones de virulencia. Los genes efectores, fueron silenciados en una cepa, para Avr-vnt1 en POX109 y para el homólogo Avh9.1 en POX067, pero expresados en la otra. Los resultados de transcriptoma fueron comparados con tres cepas adicionales del linaje EC-1. La información del repertorio de los efectores del patógeno y su expresión podrían ser informativos para el mejoramiento genético de la resistencia. El descubrimiento de efectores silenciados en las poblaciones del patógeno pueden guiar al uso de genes R específicos en los programas de mejoramiento genético. Por ejemplo, en el contexto de los Andes, donde el linaje clonal EC-1 predomina, el gen Rpi-vnt1 podría no ser recomendado.

Potato (Solanum tuberosum L.) is nowadays one of the most important food crops. It is prone to the disease known Late blight caused by the oomycete pathogen Phytophthora infestans, which secrete a hundreds of effectors that act as virulence factors. Little is known of the diversity of virulence genes of the strains that belong to a clonally reproducing lineage EC-1. In this paper, through the transcriptome sequencing of potato and P. infestans interaction, we identified differentially expressed RXLR type effector genes in two P. infestans isolates EC-1, being confirmed by qRT-PCR. The isolates, originate from cultivated potato in the central Peruvian Andes, have different virulence patterns. Effector genes, were silenced in one isolate, such as Avr-vnt1 in POX 109 and for Avh9.1 homolog in POX 067, but expressed in the other. The transcriptomics results were compared with three additional isolates from the EC-1 lineage. The information on the pathogen effector repertoire and its expression should be informative for resistance breeding. Discovery of silenced effectors in the pathogen populations can guide the use of specific R genes in the breeding programs. For example in the Andean setting where EC-1 lineage dominates the Rpi-vnt1 would not be recommended.

Aislamiento y caracterización de cepas de Bacillus spp. con actividad contra Tetranychus urticae Koch en cultivos comerciales de rosas / Isolation and characterization of strains of Bacillus spp. With activity against Tetranychus urticae Koch in commercial crops of roses

Una de las especies de ácaros que producen considerables pérdidas cualitativas y cuantitativas en el cultivo de rosas bajo invernadero en Ecuador es Tetranychus urticae, donde el control con agroquímicos convencionales no es efectivo; por lo cual se busca identificar cepas de Bacillus spp. aisladas a partir de T. urticae y determinar sus escenarios anatómicos patogénicos para un futuro control con bacterias. La fase de campo se realizó en la florícola Naranjo Roses S.A. en Latacunga en un cultivo de rosas bajo condiciones orgánicas, en donde se recolectaron hojas de cada tercio de seis plantas con presencia de T. urticae Koch. Las muestras fueron trasladadas para ser procesadas en Plantsphere Laboratories (PSL) en Quito. Se identificaron y cuantificaron los diferentes estadios de la plaga y se aislaron algunos individuos para determinar la presencia de cepas patogénicas de Bacillus spp., las cuales fueron aisladas, purificadas e identificadas. Los tratamientos de verificación patogénica fueron las cepas, agua destilada (testigo) y Bacillus thuringiensis biovar acari (testigo positivo). Los eventos patogénicos se evaluaron en hembras adultas de T. urticae mediante citohistoquímica, por medio de la cual se reportó ruptura de paredes externas, precipitación de contenido celular y malformaciones cuticulares. Se determinó que el mayor número de individuos plaga se localizan en el tercio bajo (59,4%) con mayor presencia de huevos (63,3%). Mediante análisis de componentes principales (ACP) de los tratamientos, se determinó las cepas más eficientes como Efectores Biocatalíticos (EBc©)siendo estas la PSL 104, 113, 114 y Bacillus thuringiensis biovar acari.

One of the species of mites that cause considerable qualitative and quantitative losses in roses growing under greenhouses in Ecuador is Tetranychus urticae, where conventional agrochemicals control is not effective; therefore seeks to identify strains of Bacillus spp. Isolated from T. urticae and determine their anatomical pathogenic scenarios for future control with bacteria. The field phase took place in Naranjo Roses S.A. in Latacunga in a growing roses under organic conditions, where they were collected sheets of each third of six plants with presence of T. urticae Koch. The samples were taken to be processed in Plantsphere Laboratories (PSL) in Quito. They were identified and quantified the different stages of the pest and some individuals were isolated for the presence of pathogenic strains of Bacillus spp., which were isolated, purified and identified. Check treatments of pathogenic were strains, distilled water (control) and Bacillus thuringiensis biovar acari (positive control). Pathogenic events were evaluated in adult females of T. urticae by citohistochemistry, where breakout of the outer walls, precipitation of cell contents and cuticular malformations were reported. It was determined that the largest numbers of pest individuals are located in the lower third (59.4%) with greater presence of eggs (63.3%). Using principal component analysis (PCA) of the treatments, the PSL 104, 113, 114 and Bacillus thuringiensis biovar acari was determined as the most efficient Biocatalytic Effectors (EBc©).

Establishment and optimization for TALE-TFs construction / 重庆医学

Objective To optimize the method of transcription activator‐like effector transcription factors (TALE‐TFs) con‐struction ,some improvement and adaption were made based on the traditional methods .Methods We first constructed the basic tandem fragments with different length ,including trimer ,tetramer ,pentamer and hexamer by Golden Gate cloning technique and PCR ,then the procedure with the highest efficacy was chosen to construct our TALE‐TFs .To determine the function of the TALE‐TFs ,the plasmid pminCMV with the specific binding sequence of TALE‐TFs was constructed by fragment substitution reaction (FSR) .The transcription activating function of TALE‐TFs was confirmed by the intensity of red fluorescence ,after TALE‐TFs , pEGFP‐N1 and pminCMV plasmid were co‐transfected into 293HEK cells .Results An optimized method for TALE‐TFs construc‐tion and functional assay was established .Conclusion This method can potentially be wildly used in fields that the expression of some constitutively expressed genes needs to be modified .

Perspectives in the control of infectious diseases by transgenic mosquitoes in the post-genomic era: a review

Arthropod-borne diseases caused by a variety of microorganisms such as dengue virus and malaria parasites afflict billions of people worldwide imposing major economic and social burdens. Despite many efforts, vaccines against diseases transmitted by mosquitoes, with the exception of yellow fever, are not available. Control of such infectious pathogens is mainly performed by vector management and treatment of affected individuals with drugs. However, the numbers of insecticide-resistant insects and drug-resistant parasites are increasing. Therefore, inspired in recent years by a lot of new data produced by genomics and post-genomics research, several scientific groups have been working on different strategies to control infectious arthropod-borne diseases. This review focuses on recent advances and perspectives towards construction of transgenic mosquitoes refractory to malaria parasites and dengue virus transmission.

Establishment of FSP27 Gene Knockdown Preadipocyte Cell Line / 生物化学与生物物理进展

Obesity and its related metabolic diseases become major health problems in the world.Adipose tissue plays an important role in the development of obesity.FSP27,a member of the CIDE family proteins,is expressed at high levels in white adipose tissue and differentiated 3T3L1 cells.The objective of current study is to establish a FSP27 knockdown preadipocyte cell line to investigate the in vivo function of mouse FSP27.The double strand siRNA of mouse FSP27 corresponding to nucleotides 270 to 291 was synthesized and inserted into pSilencer2.1.pSilencer-siFSP27 was co-transfected into 293T cells with the HA-mFSP27 expression vector to test its knock-down efficiency.The FSP27 siRNA was then transferred to a lentiviral vector.Lentivirus were generated and used to infect 3T3-L1 cells.It was shown here that lentivirus containing FSP27siRNA can effectively knockdown FSP27 expression in 3T3-L1 cells.Establishment of FSP27 knock-down cell line provides a useful tool for the study of in vivo function FSP27.

The cloning and construction of eukaryotic expression vector of human CIDE3 and its proapoptotic effect on HeLa cells / 医学研究生学报

Objective:To clone the human cell death-inducing DFF45-like effectors 3(CIDE3) full length cDNA for construction the eukaryotic expression vector pcDNA3.l(+)-CIDE3 and to study its bioactivity.Methods:① Total RNA was extracted from human white adipose tissues and the desired cDNA fragment was obtained by RT-PCR.After the fragment had being inserted into an eukaryotic expression vector pcDNA3.l(+),the resulted recombinant plasmid pcDNA3.l(+)-CIDE3 was transformed into DH5?.The positive clone was selected and confirmed to contain full length of CIDE3 cDNA by agarose gel and DNA sequence analysis.②After the pcDNA3.l(+)-CIDE3 plasmid was transfected into HeLa cells under lipofectamine mediation,the effect of target gene expression on growth of HeLa cells was analysed by TUNEL staining. Results:① The recombinant eukaryotic expression plasmid pcDNA3.l(+)-CIDE3 was constructed successfully and the sequence of CIDE3 was consistent with that of genebank.②After transfected pcDNA3.l(+)-CIDE3,HeLa cells presented distinguished apoptosis(about 15%),compared with that of transfected plasmid pcDNA3.l(+)(

Regulation of cellular signals by G-proteins.

Extracellular signals are transduced across the cell by the cell surface receptors, with the aid of G-proteins, which act at a critical point of signal transduction and cellular regulation. Structurally, G-proteins are heterotrimeric consisting α, β and γ subunits but in functionally active state they dissociate into α subunit coupled to GTP and as βγ dimer. G-proteins can be broadly divided into two classes based on their sensitivity to pertussis toxin and cholera toxin. Existence of various forms of each of the subunit allows molecular diversity in the subunit species of G-proteins. These subunits interact with a wide range of receptors and effectors, facilitated by post translational modification of their subunits. Different types of G-proteins mediate several signalling events in different parts of the body. This review summarizes the features of (i) structural and functional heterogenity among different subunits of G-proteins, (ii) interaction of G-proteins and their subunits with effectors with specific cases of G-protein mediated signalling in olfaction, phototransduction in the retina, ras and ras related transduction and (iii) disease conditions associated with malfunctioning of G-proteins.