Application of scanning electron microscopy for diagnosing phytoplasmas in single and mixed (virus-phytoplasma ) infection in Papaya

Phytoplasma and some viruses, papaya ring spot (PRSV) and papaya mosaic (PapMV) have been reported in papaya, from different Mexican states. Some symptoms of yellow type diseases, such as mosaics, stunting, bunchy top and leaf chlorosis, necrosis and malformations are somewhat similar in appearance, but caused by distinct pathogens. Using a scanning electron microscopy (SEM) technique phytoplasmas were detected in the phloem tissues of field and greenhouse-indexed papaya plants from Baja California Sur (BCS). Samples from 32 local varieties, as well as cv. Maradol, showing numerous symptoms of dieback, mosaics, bunchy top, and yellow crinkle were analyzed. The pathogen was detected in stems, leafstalks, roots, axillary leaflets, leaf veins and flowers. Phytoplasma was also detected in dry and in germinated seeds within the fruit, suggesting seed transmission of the pathogen. Some ultrastructural peculiarities of phytoplasma in infected tissues were also observed. No viral infection with PRSV and PapMV was revealed neither in test-plants nor by molecular techniques. Application of SEM technique for analysis of papaya samples from Veracruz and Irapuato, both from field-grown and mechanically inoculated plants with PRSV and PapMV in various combinations also revealed phytoplasmas in the phloem of most of tested samples. In some cases, along with phytoplasmas, rod-shaped bacteria were distinguished.Keywords: Papaya, phytoplasma, papaya ringspot virus, papaya mosaic virus, scanning electron microscopy, Mexic

Can Children Have Ordinary Expectable Caregiving Environments in Unconventional Contexts? Quality of Care Organization in Three Mexican Same-Sex Planned Families

The aim of this research was to explore the elements that configure the quality of care among three Mexican same-sex planned families: two female-parented families (through donor insemination) and a male-parented one (through adoption). The first family consisted of two mothers and a 3-year-old daughter; the second one had two mothers and a 1.5-year-old set of boy twins and the third family consisted of two fathers and a 2-year-old girl. It was assumed that Ainsworth’s notions of quality of care organization are useful in order to understand caregiver–child attachment relationships, regardless of the parents’ sexual orientation. A collective case study was selected due to the fact that these families shared their “unconventionality” (i.e., parents were not heterosexual) and the fact that they were planned, but each one constituted a particular case with a unique configuration. Four trained independent observers used the q-sort methodology (Maternal Behavior Q-Sort and Attachment Q-Sort) to describe parents’ and children’s behavior, respectively. The findings showed that parents were highly sensitive and all children used them as a secure base. To provide an in-depth examination of which elements configure the quality of care, a semi-structured interview with each parent was carried out. Through a thematic analysis, an over-arching theme named Affections and Emotions was identified, together with six subthemes: (1) Creating an affective environment; (2) Being available; (3) Acknowledging and expressing emotions; (4) Perceiving, interpreting and responding adequately to the child’s real self; (5) Taking the child’s perspective into account; and (6) Agreeing on roles and dividing the tasks. In order to showcase the particular configuration of gay parenting, the male-headed family narrative is reported in detail, because gay parents have been perceived as violating traditional gender roles as well as the hegemonic model of masculinity. The findings were consistent with the notion of quality of care as proposed by Ainsworth and her collaborators. The implications of the methodological device and research regarding same-sex planned families are discussed so as to understand the organization of the caregiving environment

Desarrollo y crisis en el capitalismo

Revisión realizada en 2012, del desenvolvimiento de la economía mundial y los cambios que se han venido dando en el contexto de la crisis reciente, así como diversos aspectos de las transformaciones en proceso y de la situación económica y social en América Latina. La primera parte está dedicada a la crisis mundial en el marco del funcionamiento del capitalismo y la globalización, y de la permanencia de los factores económicos y financieros que propiciaron su estallido. Se consideran los efectos de la misma según los países y regiones, y se observa su incidencia en los procesos de reconfiguración en marcha, así como las estrategias y políticas seguidas para hacerle frente, sin olvidar las propuestas que se han adelantado para explicarla. En la segunda parte de la obra se analizan los problemas de la región latinoamericana y los de algunos de los países que la integran. Los temas incluyen el desempleo juvenil y el modelo agro-exportador, la inversión extranjera directa y las elecciones presidenciales, entre otros

Peripheral T-lymphocytes express WNT7A and its restoration in leukemia-derived lymphoblasts inhibits cell proliferation

<p>Abstract</p> <p>Background</p> <p>WNT7a, a member of the Wnt ligand family implicated in several developmental processes, has also been reported to be dysregulated in some types of tumors; however, its function and implication in oncogenesis is poorly understood. Moreover, the expression of this gene and the role that it plays in the biology of blood cells remains unclear. In addition to determining the expression of the <it>WNT7A </it>gene in blood cells, in leukemia-derived cell lines, and in samples of patients with leukemia, the aim of this study was to seek the effect of this gene in proliferation.</p> <p>Methods</p> <p>We analyzed peripheral blood mononuclear cells, sorted CD3 and CD19 cells, four leukemia-derived cell lines, and blood samples from 14 patients with Acute lymphoblastic leukemia (ALL), and 19 clinically healthy subjects. Reverse transcription followed by quantitative Real-time Polymerase chain reaction (qRT-PCR) analysis were performed to determine relative <it>WNT7A </it>expression. Restoration of WNT7a was done employing a lentiviral system and by using a recombinant human protein. Cell proliferation was measured by addition of WST-1 to cell cultures.</p> <p>Results</p> <p>WNT7a is mainly produced by CD3 T-lymphocytes, its expression decreases upon activation, and it is severely reduced in leukemia-derived cell lines, as well as in the blood samples of patients with ALL when compared with healthy controls (<it>p </it>≤0.001). By restoring <it>WNT7A </it>expression in leukemia-derived cells, we were able to demonstrate that WNT7a inhibits cell growth. A similar effect was observed when a recombinant human WNT7a protein was used. Interestingly, restoration of <it>WNT7A </it>expression in Jurkat cells did not activate the canonical Wnt/β-catenin pathway.</p> <p>Conclusions</p> <p>To our knowledge, this is the first report evidencing quantitatively decreased <it>WNT7A </it>levels in leukemia-derived cells and that <it>WNT7A </it>restoration in T-lymphocytes inhibits cell proliferation. In addition, our results also support the possible function of <it>WNT7A </it>as a tumor suppressor gene as well as a therapeutic tool.</p

Cutting Edge:Transcriptional profiling reveals multifunctional and cytotoxic antiviral responses of zika virus-specific CD8 + T cells

Zika virus (ZIKV) constitutes an increasing public health problem. Previous studies have shown that CD8 + T cells play an important role in ZIKV-specific protective immunity. We have previously defined antigenic targets of the ZIKV-specific CD8 + T cell response in humans. In this study, we characterized the quality and phenotypes of these responses by a combined use of flow cytometry and transcriptomic methods, using PBMCs from donors deriving from different geographical locations collected in the convalescent phase of infection. We show that ZIKVspecific CD8 + T cells are characterized by a polyfunctional IFN-g signature with upregulation of TNF-a, TNF receptors, and related activation markers, such as CD69, as well as a cytotoxic signature characterized by strong upregulation of GZMB and CRTAM. The signature is stable and not influenced by previous dengue virus exposure, geographical location, or time of sample collection postinfection. To our knowledge, this work elucidates the first in-depth characterization of human CD8 + T cells responding to ZIKV infection

Vermiculite bio-barriers for Cu and Zn remediation: an eco-friendly approach for freshwater and sediments protection

The increase in heavy metal contamination in freshwater systems causes serious environmental problems in most industrialized countries, and the effort to find ecofriendly techniques for reducing water and sediment contamination is fundamental for environmental protection. Permeable barriers made of natural clays can be used as low-cost and eco-friendly materials for adsorbing heavy metals from water solution and thus reducing the sediment contamination. This study discusses the application of permeable barriers made of vermiculite clay for heavy metals remediation at the interface between water and sediments and investigates the possibility to increase their efficiency by loading the vermiculite surface with a microbial biofilm of Pseudomonas putida, which is well known to be a heavy metal accumulator. Some batch assays were performed to verify the uptake capacity of two systems and their adsorption kinetics, and the results indicated that the vermiculite bio-barrier system had a higher removal capacity than the vermiculite barrier (?34.4 and 22.8 % for Cu and Zn, respectively). Moreover, the presence of P. putida biofilm strongly contributed to fasten the kinetics of metals adsorption onto vermiculite sheets. In open-system conditions, the presence of a vermiculite barrier at the interface between water and sediment could reduce the sediment contamination up to 20 and 23 % for Cu and Zn, respectively, highlighting the efficiency of these eco-friendly materials for environmental applications. Nevertheless, the contribution of microbial biofilm in open-system setup should be optimized, and some important considerations about biofilm attachment in a continuous-flow system have been discussed.This work has been produced thanks to the collaboration of Dip.SA (University of Bologna) and IBB (University of Minho). A particular acknowledgment is due to Dr. E. Rosales. The work was partially financed by the FCT Strategic Project Pest-OE/EQB/LA0023 and the Project ‘‘BioEnv—Biotechnology and Bioengineering for a sustainable world,’’ co-funded by the Programa Operacional Regional do Norte (ON.2–O Novo Norte), QREN, FEDER

Patients with primary immunodeficiencies are a reservoir of poliovirus and a risk to polio eradication

ABSTARCT: Immunodeficiency-associated vaccine-derived polioviruses (iVDPVs) have been isolated from primary immunodeficiency (PID) patients exposed to oral poliovirus vaccine (OPV). Patients may excrete poliovirus strains for months or years; the excreted viruses are frequently highly divergent from the parental OPV and have been shown to be as neurovirulent as wild virus. Thus, these patients represent a potential reservoir for transmission of neurovirulent polioviruses in the post-eradication era. In support of WHO recommendations to better estimate the prevalence of poliovirus excreters among PIDs and characterize genetic evolution of these strains, 635 patients including 570 with primary antibody deficiencies and 65 combined immunodeficiencies were studied from 13 OPV-using countries. Two stool samples were collected over 4 days, tested for enterovirus, and the poliovirus positive samples were sequenced. Thirteen patients (2%) excreted polioviruses, most for less than 2 months following identification of infection. Five (0.8%) were classified as iVDPVs (only in combined immunodeficiencies and mostly poliovirus serotype 2). Non-polio enteroviruses were detected in 30 patients (4.7%). Patients with combined immunodeficiencies had increased risk of delayed poliovirus clearance compared to primary antibody deficiencies. Usually, iVDPV was detected in subjects with combined immunodeficiencies in a short period of time after OPV exposure, most for less than 6 months. Surveillance for poliovirus excretion among PID patients should be reinforced until polio eradication is certified and the use of OPV is stopped. Survival rates among PID patients are improving in lower and middle income countries, and iVDPV excreters are identified more frequently. Antivirals or enhanced immunotherapies presently in development represent the only potential means to manage the treatment of prolonged excreters and the risk they present to the polio endgame. Keywords: Poliovirus eradication, Immunodeficiency-associated vaccine-derived polioviruses, Oral poliovirus vaccine, Humoral immunodeficiency, Combined immunodeficiency, Primary immunodeficienc

Characterization of the genetic diversity of Mycobacterium tuberculosis in São Paulo city, Brazil

<p>Abstract</p> <p>Background</p> <p>Tuberculosis is a major health problem in São Paulo, Brazil, which is the most populous and one of the most cosmopolitan cities in South America. To characterize the genetic diversity of <it>Mycobacterium tuberculosis </it>in the population of this city, the genotyping techniques of spoligotyping and MIRU were applied to 93 isolates collected in two consecutive years from 93 different tuberculosis patients residing in São Paulo city and attending the Clemente Ferreira Institute (the reference clinic for the treatment of tuberculosis).</p> <p>Findings</p> <p>Spoligotyping generated 53 different spoligotype patterns. Fifty-one isolates (54.8%) were grouped into 13 spoligotyping clusters. Seventy- two strains (77.4%) showed spoligotypes described in the international databases (SpolDB4, SITVIT), and 21 (22.6%) showed unidentified patterns. The most frequent spoligotype families were Latin American Mediterranean (LAM) (26 isolates), followed by the T family (24 isolates) and Haarlem (H) (11 isolates), which together accounted for 65.4% of all the isolates. These three families represent the major genotypes found in Africa, Central America, South America and Europe. Six Spoligo-International-types (designated SITs by the database) comprised 51.8% (37/72) of all the identified spoligotypes (SIT53, SIT50, SIT42, SIT60, SIT17 and SIT1). Other SITs found in this study indicated the great genetic diversity of <it>M. tuberculosis</it>, reflecting the remarkable ethnic diversity of São Paulo city inhabitants. The MIRU technique was more discriminatory and did not identify any genetic clusters with 100% similarity among the 93 isolates. The allelic analysis showed that MIRU loci 26, 40, 23 and 10 were the most discriminatory. When MIRU and spoligotyping techniques were combined, all isolates grouped in the 13 spoligotyping clusters were separated.</p> <p>Conclusions</p> <p>Our data indicated the genomic stability of over 50% of spoligotypes identified in São Paulo and the great genetic diversity of <it>M. tuberculosis </it>isolates in the remaining SITs, reflecting the large ethnic mix of the São Paulo city inhabitants. The results also indicated that in this city, <it>M. tuberculosis </it>isolates acquired drug resistance independently of genotype and that resistance was more dependent on the selective pressure of treatment failure and the environmental circumstances of patients.</p