ABSTRACT
Objetivo: descripción de hallazgos básicos para establecer sospecha de malformación cloacal. Material y métodos: estudio descriptivo de caso único de malformación cloacal, sospechado tras repetición de ecografía de primer trimestre por traslucencia nucal > p-99, con sacos linfáticos yugulares bilaterales y cribado combinado de 1/968. Resultados: presentamos el caso de una gestante cuya sospecha se estableció en la semana 15 de gestación mediante control ecográfico seriado y con diagnóstico anatomo-patológico definitivo tras estudio necrópsico al realizar interrupción legal del embarazo. Conclusiones: hemos sospechado esta rara y severa entidad precozmente mediante signos indirectos que induce a un examen exhaustivo. De esta manera, es posible llegar a su diagnóstico definitivo y clasificar su severidad, futuro pronóstico y posible tratamiento (AU)
Objective: Description of basic findings to be able to suspect cloacal malformation. Material and methods: Descriptive study of a unique case of cloacal malformation, suspected in a repeated ultrasound of the first trimester because of a nucal translucency of > p-99, with bilateral yugular saccules and combined risk 1/968. Results: We report the case of a pregnant woman, in which we suspected this malformation during de 15th week of gestation with repeated ultrasound, and with a definitive anatomy- pathological diagnosis as she performed a legal interruption of de gestation. Conclusion: We have suspected this strange and severe entity in a early time because of indirect signs that obligated us to an exhaustive exam. In this way, it is possible to establish a definitive diagnose and severity, future prognosis and treatment possibilities (AU)
Subject(s)
Humans , Female , Pregnancy , Adult , Cloaca/abnormalities , Urogenital Abnormalities/diagnostic imaging , Gastrointestinal Tract/abnormalities , Ultrasonography, Prenatal , Abortion, Legal , Nuchal Translucency Measurement , Amniocentesis , Diagnosis, DifferentialABSTRACT
In the Potyvirus genus, the P1 protein is the first N-terminal product processed from the viral polyprotein, followed by the helper-component proteinase (HCPro). In silencing suppression patch assays, we found that Potato virus Y (PVY) HCPro expressed from a P1-HCPro sequence increased the accumulation of a reporter gene, whereas protein expressed from an HCPro sequence did not, even with P1 supplied in trans. This enhancing effect of P1 has been noted in other potyviruses, but has remained unexplained. We analysed the accumulation of PVY HCPro in infiltrated tissues and found that it was higher when expressed from P1-HCPro than from HCPro sequences. Co-expression of heterologous suppressors increased the steady-state level of mRNA expressed from the HCPro sequence, but not that of protein. This suggests that, in the absence of P1 upstream, either HCPro acquires a conformation that affects negatively its activity or stability, or that its translation is reduced. To test these options, we purified HCPro expressed in the presence or absence of upstream P1, and found no difference in purification pattern and final soluble state. By contrast, alteration of the Kozak context in the HCProâ mRNA sequence to favour translation increased partially suppressor accumulation and activity. Furthermore, protein activity was not lower than in protein expressed from P1-HCPro sequences. Thus, a direct role for P1 on HCPro suppressor activity or stability, by influencing its conformation during translation, can be excluded. However, P1 could still have an indirect effect favouring HCPro accumulation. Our data highlight the relevance of cis-acting translational elements in the heterologous expression of HCPro.
Subject(s)
Cysteine Endopeptidases/metabolism , Gene Silencing , Potyvirus/metabolism , Protein Biosynthesis , Viral Proteins/metabolism , Amino Acid Sequence , Base Sequence , Cysteine Endopeptidases/chemistry , Cysteine Endopeptidases/isolation & purification , Genes, Suppressor , Molecular Sequence Data , Potyvirus/genetics , Solubility , Suppression, Genetic , Nicotiana/virology , Viral Proteins/chemistry , Viral Proteins/isolation & purificationABSTRACT
Groundnut bud necrosis virus (GBNV) infects a large number of leguminous and solanaceous plants. To elucidate the biological function of the non-structural protein encoded by the S RNA of GBNV (NSs), we studied its role in RNA silencing suppression and in viral pathogenesis. Our results demonstrated that GBNV NSs functions as a suppressor of RNA silencing using the agroinfiltration patch assay. An in silico analysis suggested the presence of pro-apoptotic protein Reaper-like sequences in the GBNV NSs, which were known to be present in animal infecting bunyaviruses. Utilizing NSs mutants, we demonstrated that a Leu-rich domain was required for RNA silencing suppression activity, but not the non-overlapping Trp/GH3 motif of the Reaper-like sequence. To investigate the role of NSs in symptom development we generated transgenic tomato expressing the GBNV NSs and showed that the expression of NSs in tomato mimics symptoms induced by infection with GBNV, such as leaf senescence and necrosis. As leaf senescence is controlled by miR319 regulation of the transcription factor TCP1, we assessed the accumulation of both RNAs in transgenic NSs-expressing and GBNV-infected tomato plants. In both types of plants the levels of miR319 decreased, while the levels of TCP1 transcripts increased. We propose that GBNV-NSs affects miRNA biogenesis through its RNA silencing suppressor activity and interferes with TCP1-regulated leaf developmental pathways.
