Induction of Monoterpenoid Oxindole Alkaloids Production and Related Biosynthetic Gene Expression in Response to Signaling Molecules in Hamelia patens Plant Cultures.

Hamelia patens (Rubiaceae), known as firebush, is a source of bioactive monoterpenoid oxindole alkaloids (MOAs) derived from monoterpenoid indole alkaloids (MIAs). With the aim of understanding the regulation of the biosynthesis of these specialized metabolites, micropropagated plants were elicited with jasmonic acid (JA) and salicylic acid (SA). The MOA production and MIA biosynthetic-related gene expression were evaluated over time. The production of MOAs was increased compared to the control up to 2-fold (41.3 mg g DW-1) at 72 h in JA-elicited plants and 2.5-fold (42.4 mg g DW-1) at 120 h in plants elicited with SA. The increment concurs with the increase in the expression levels of the genes HpaLAMT, HpaTDC, HpaSTR, HpaNPF2.9, HpaTHAS1, and HpaTHAS2. Interestingly, it was found that HpaSGD was downregulated in both treatments after 24 h but in the SA treatment at 120 h only was upregulated to 8-fold compared to the control. In this work, we present the results of MOA production in H. patens and discuss how JA and SA might be regulating the central biosynthetic steps that involve HpaSGD and HpaTHAS genes.

Whole-Exome Sequencing of 21 Families: Candidate Genes for Early-Onset High Myopia.

High myopia is the most severe and pathological form of myopia. It occurs when the spherical refractive error exceeds -6.00 spherical diopters (SDs) or the axial length (AL) of the eye is greater than 26 mm. This article focuses on early-onset high myopia, an increasingly common condition that affects children under 10 years of age and can lead to other serious ocular pathologies. Through the genetic analysis of 21 families with early-onset high myopia, this study seeks to contribute to a better understanding of the role of genetics in this disease and to propose candidate genes. Whole-exome sequencing studies with a panel of genes known to be involved in the pathology were performed in families with inconclusive results: 3% of the variants found were classified as pathogenic, 6% were likely pathogenic and the remaining 91% were variants of uncertain significance. Most of the families in this study were found to have alterations in several of the proposed genes. This suggests a polygenic inheritance of the pathology due to the cumulative effect of the alterations. Further studies are needed to validate and confirm the role of these alterations in the development of early-onset high myopia and its polygenic inheritance.

Protein S-acylation controls the subcellular localization and biological activity of PHYTOCHROME KINASE SUBSTRATE.

PHYTOCHROME KINASE SUBSTRATE (PKS) proteins are involved in light-modulated changes in growth orientation. They act downstream of phytochromes to control hypocotyl gravitropism in the light and act early in phototropin signaling. Despite their importance for plant development, little is known about their molecular mode of action, except that they belong to a protein complex comprising phototropins at the plasma membrane (PM). Identifying evolutionary conservation is one approach to revealing biologically important protein motifs. Here, we show that PKS sequences are restricted to seed plants and that these proteins share 6 motifs (A to F from the N to the C terminus). Motifs A and D are also present in BIG GRAIN, while the remaining 4 are specific to PKSs. We provide evidence that motif C is S-acylated on highly conserved cysteines, which mediates the association of PKS proteins with the PM. Motif C is also required for PKS4-mediated phototropism and light-regulated hypocotyl gravitropism. Finally, our data suggest that the mode of PKS4 association with the PM is important for its biological activity. Our work, therefore, identifies conserved cysteines contributing to PM association of PKS proteins and strongly suggests that this is their site of action to modulate environmentally regulated organ positioning.

Acute Fibrinous Anterior Uveitis in a Patient Treated With Topical Interferon-α2b for Ocular Surface Squamous Neoplasia With Uveitis Susceptibility.

PURPOSE: To report a case of fibrinous acute anterior uveitis associated with topical interferon-α2b (IFN-α2b) treatment for ocular surface squamous neoplasia in a patient with HLA-B27 uveitis predisposition. METHODS: Case report. RESULTS: We present the case of a 57-year-old man who received topical IFN-α2b as adjuvant therapy for a previously surgically removed ocular surface squamous neoplasia with affected surgical margins. Two weeks after topical IFN-α2b initiation, the patient was diagnosed with fibrinous acute anterior uveitis. Complementary tests to rule out other causes of uveitis resulted to be negative, except for HLA-B27, which tested positive. Response to treatment with topical corticosteroids and cyclopentolate was favorable. As IFN-α2b is considered an immune enhancer and has been widely associated with autoimmune side effects, topical therapy with IFN-α2b was temporally ceased until intraocular inflammation resolved. Topical IFN-α2b was resumed, and during follow-up, no signs of uveitis were detected. The main hypothesis is that IFN-α2b acts as a trigger for intraocular inflammation in individuals with uveitis predisposition. CONCLUSIONS: Topical IFN-α2b could trigger intraocular inflammation in patients with uveitis susceptibility. It may be reasonable to use IFN-α2b cautiously in patients with a known history of uveitis or uveitis predisposition.

Next-Generation Sequencing Screening of 43 Families with Non-Syndromic Early-Onset High Myopia: A Clinical and Genetic Study.

Early-onset high myopia (EoHM) is a disease that causes a spherical refraction error of ≥-6 diopters before 10 years of age, with potential multiple ocular complications. In this article, we report a clinical and genetic study of 43 families with EoHM recruited in our center. A complete ophthalmological evaluation was performed, and a sample of peripheral blood was obtained from proband and family members. DNA was analyzed using a customized next-generation sequencing panel that included 419 genes related to ophthalmological disorders with a suspected genetic cause, and genes related to EoHM pathogenesis. We detected pathogenic and likely pathogenic variants in 23.9% of the families and detected variants of unknown significance in 76.1%. Of these, 5.7% were found in genes related to non-syndromic EoHM, 48.6% in genes associated with inherited retinal dystrophies that can include a syndromic phenotype, and 45.7% in genes that are not directly related to EoHM or retinal dystrophy. We found no candidate genes in 23% of the patients, which suggests that further studies are needed. We propose a systematic genetic analysis for patients with EoHM because it helps with follow-up, prognosis and genetic counseling.

Respuesta clínica y neurofisiológica a la efedrina en un paciente con síndrome miasténico congénito de canal lento / Clinical and neurophysiological response to ephedrine in a patient affected with slow-channel congenital myasthenic syndrome

INTRODUCCIÓN: El síndrome miasténico congénito de canal lento, o síndrome de canales lentos, es un trastorno neuromuscular progresivo hereditario, autosómico dominante, causado por una activación anormal de los receptores de la acetilcolina en la unión neuromuscular. La alteración histopatológica característica es la degeneración selectiva de la placa terminal y la membrana postsináptica debido a la sobrecarga de calcio. La piridostigmina debe evitarse en este síndrome, y la quinidina o la fluoxetina son las terapias recomendadas actualmente. CASO CLÍNICO: Niña de 11 años con un fenotipo de cinturas de síndrome miasténico congénito de canal lento que presenta debilidad y fatiga lentamente progresivas desde los 8 años. Tras un empeoramiento clínico con piridostigmina, iniciado empíricamente antes de que los resultados de la secuenciación del exoma estuvieran disponibles, se observó una respuesta espectacular y sostenida con efedrina en monoterapia. La secuenciación del exoma reveló una mutación heterocigota de novo en el gen CHRNB1: c.865G>A; p.Val289Met (NM_000747.2). El estudio electromiográfico con estimulación repetitiva en el nervio peroneo mostró una disminución anormal en la amplitud (23,9%) y también la génesis de un segundo potencial de acción muscular compuesto más pequeño después del pico de la onda M principal en los nervios motores mediano, cubital y peroneo. CONCLUSIÓN: Aunque se han documentado respuestas favorables a agonistas adrenérgicos en asociación con la fluoxetina, ésta representa la primera aportación que documenta una respuesta clínica relevante con efedrina en monoterapia en un paciente con síndrome miasténico congénito de canal lento. Los agonistas adrenérgicos pueden considerarse una opción terapéutica en pacientes con este síndrome

INTRODUCTION: Slow-channel congenital myasthenic syndrome is an autosomal dominant inherited progressive neuromuscular disorder caused by abnormal gating of mutant acetylcholine receptors in the neuromuscular junction. Its pathological hallmark is selective degeneration of the endplate and postsynaptic membrane due to calcium overload. Pyridostigmine should be avoided in this syndrome, being quinidine or fluoxetine the current recommended therapies. CASE REPORT: An 11-year-old girl with a limb-girdle phenotype of slow-channel congenital myasthenic syndrome presenting with a slowly progressive fatigable weakness at the age of 8 years. After a clinical worsening with pyridostigmine, empirically started before the exome sequencing results were available, a dramatic and sustained response to ephedrine monotherapy was observed. Whole exome sequencing revealed a de novo heterozygous mutation in CHRNB1 gene: c.865G>A; p.Val289Met (NM_000747.2). An abnormal decrement in amplitude (23.9%) from the first to fifth intravollley waveform was revealed after repetitive peroneal nerve stimulation at low frequencies. In addition, a second smaller compound muscle action potential after the peak of the main M-wave in median, ulnar and peroneal motor nerves was observed. CONCLUSION: Favorable responses to adrenergic agonists added to fluoxetine had been reported. However, to the best of our knowledge this is the first report on effective monotherapy with ephedrine in a slow-channel congenital myasthenic syndrome patient. Adrenergic agonists may be considered as a therapeutic option in patients with this syndrome

Mutación de novo en heterocigosis en el gen MBD5 asociada a heterotopía en banda bilateral y polimicrogiria / De novo heterozygous mutation in the MBD5 gene associated with bilateral band heterotopia and polymicrogyria

Introducción: La etiología del retraso mental autosómico dominante 1, también conocido como pseudo-Angelman, trastorno del neurodesarrollo asociado a MBD5 o haploinsuficiencia MBD5, radica en una microdeleción del cromosoma 2q23.1 o en una alteración específica del gen MBD5, que constituye la mínima región afectada en la citada microdeleción. Objetivo: Comunicar el caso de una niña con una mutación heterocigota y de novo en el gen MBD5 asociada a heterotopía en banda bilateral y polimicrogiria. Caso clínico: Niña de 8 años, seguida evolutivamente desde los 18 meses por presentar la asociación de discapacidad intelectual y retraso motor graves, ausencia de desarrollo del lenguaje, hipotonía segmentaria, frente ancha y cifoescoliosis. En la resonancia magnética cerebral se observó la presencia de una heterotopía en banda bilateral y polimicrogiria parietooccipital de predominio izquierdo. En el exoma se detectó la variante de novo c.397+1G > C en heterocigosis en el gen MBD5. Conclusión: Constituye la primera observación con una mutación heterocigota en el gen MBD5 asociada a un trastorno en la migración neuronal

Introduction: The aetiology of autosomal dominant mental retardation type 1, also known as pseudo-Angelman, MBD5-associated neurodevelopmental disorder or MBD5 haploinsufficiency, lies in a microdeletion of chromosome 2q23.1 or in a specific alteration of the MBD5 gene, which constitutes the minimum region affected in the aforementioned microdeletion. Aim: To report the case of a girl with a heterozygous de novo mutation in the MBD5 gene associated with bilateral band heterotopia and polymicrogyria. Case Report: We report the case of an 8-year-old girl who was submitted to a developmental follow-up from the age of 18 months after presenting the association of severe intellectual disability and motor delay, lack of language development, segmental hypotonia, a wide forehead and kyphoscoliosis. Magnetic resonance imaging of the brain revealed the presence of a bilateral band heterotopia and parietooccipital polymicrogiria predominant on the left side. In the exome the de novo heterozygous variant c.397+1G > C was detected in the MBD5 gene. Conclusion: This is the first observation of a heterozygous mutation in the MBD5 gene associated with a neuronal migration disorder

Incremento en el número de casos diagnosticados de tosferina; ¿pensamos más en ella? / Increase in the number of diagnosed cases of whooping cough; Do we think more in it?

No disponible

Información a los padres sobre la profilaxis de la enfermedad hemorrágica del recién nacido / Information to parents on prophylactic treatment of haemorrhagic disease of the newborn

No disponible

Análisis de la utilización del dímero D en urgencias: ajuste por edad, uso inapropiado y predicción de extensión y gravedad de la embolia pulmonar / D-dimer testing in the emergency department: age adjustment, inappropriateuse, and ability to predict the extension and severity of pulmonary embolism

Objetivos: Valorar la mejora de la precisión diagnóstica del dímero D (DD) al ajustar por la edad aplicando la fórmula publicada por Douma et al., así como evaluar la adecuación de la solicitud del DD a la sospecha clínica y relacionar sus valores con la extensión y gravedad de la embolia pulmonar. Método: Estudio observacional, retrospectivo que incluye 1.833 pacientes atendidos en el servicio de urgencias de nuestro hospital a lo largo de 1 año, a los que se solicitó determinación del DD. Se calculó sensibilidad, especificidad, valor predictivo positivo y negativo de su valor utilizando el punto de corte de nuestro centro (250 μg/mL) y ajustado por edad (fórmula de Douma et al. modificada) y se correlacionó con la extensión y gravedad de la embolia pulmonar cuando la hubo. Resultados: El ajuste por edad del valor de DD supone aumentar la proporción de casos verdaderos negativos, la especificidad y el valor predictivo positivo de esta determinación. Se encuentra una correlación significativa entre el valor del DD con la extensión de la embolia pulmonar (r = 0,41, p < 0,05) pero no con la gravedad clínica del episodio. Conclusiones: El ajuste del DD por la edad mejora la precisión diagnóstica de la embolia pulmonar, aunque, en nuestro entorno, su sospecha no se establece siguiendo las guías recomendadas. El valor del DD se relaciona con la extensión pero no con la gravedad de la embolia pulmonar (AU)

Objectives: To evaluate whether using D-dimer test results adjusted for age according to the formula proposed by Douma et al. improves diagnostic accuracy; to assess the appropriateness of ordering D-dimer tests on clinical suspicion of pulmonary embolism; and to explore the association of test results with the extension and severity of the embolism. Methods: Retrospective observational study of 1833 cases in which D-dimer testing was ordered for patients in our hospital’s emergency department in the course of a year. We calculated sensitivity, specificity, and positive and negative predictive values using our hospital’s D-dimer cutoff of 250 μg/mL adjusted for age with a modification of Douma et al.’s formula. When information about pulmonary embolism extension and severity was on record, we assessed the correlation with test results. Results: Adjusting D-dimer level for age increased the number of true negatives and the specificity and positive predictive value of the test. D-dimer level correlated significantly with the extension of pulmonary embolism (r=0.41, P<.05) but not with clinical severity. Conclusions: Adjusting the D-dimer test result by age improves accuracy in the diagnosis of pulmonary embolism, even though clinical suspicion in Spain does not follow guideline recommendations. Our findings suggest that Ddimer level correlates with the extension but not the severity of pulmonary embolism (AU)

[D-dimer testing in the emergency department: age adjustment, inappropriate use, and ability to predict the extension and severity of pulmonary embolism]. / Análisis de la utilización del dímero D en urgencias: ajuste por edad, uso inapropiado y predicción de extensión y gravedad de la embolia pulmonar.

OBJECTIVES: To evaluate whether using D-dimer test results adjusted for age according to the formula proposed by Douma et al. improves diagnostic accuracy; to assess the appropriateness of ordering D-dimer tests on clinical suspicion of pulmonary embolism; and to explore the association of test results with the extension and severity of the embolism. MATERIAL AND METHODS: Retrospective observational study of 1833 cases in which D-dimer testing was ordered for patients in our hospital's emergency department in the course of a year. We calculated sensitivity, specificity, and positive and negative predictive values using our hospital's D-dimer cutoff of 250 µg/mL adjusted for age with a modification of Douma et al.'s formula. When information about pulmonary embolism extension and severity was on record, we assessed the correlation with test results. RESULTS: Adjusting D-dimer level for age increased the number of true negatives and the specificity and positive predictive value of the test. D-dimer level correlated significantly with the extension of pulmonary embolism (r=0.41, P<.05) but not with clinical severity. CONCLUSION: Adjusting the D-dimer test result by age improves accuracy in the diagnosis of pulmonary embolism, even though clinical suspicion in Spain does not follow guideline recommendations. Our findings suggest that Ddimer level correlates with the extension but not the severity of pulmonary embolism.

OBJETIVO: Valorar la mejora de la precisión diagnóstica del dímero D (DD) al ajustar por la edad aplicando la fórmula publicada por Douma et al., así como evaluar la adecuación de la solicitud del DD a la sospecha clínica y relacionar sus valores con la extensión y gravedad de la embolia pulmonar. METODO: Estudio observacional, retrospectivo que incluye 1.833 pacientes atendidos en el servicio de urgencias de nuestro hospital a lo largo de 1 año, a los que se solicitó determinación del DD. Se calculó sensibilidad, especificidad, valor predictivo positivo y negativo de su valor utilizando el punto de corte de nuestro centro (250 µg/mL) y ajustado por edad (fórmula de Douma et al. modificada) y se correlacionó con la extensión y gravedad de la embolia pulmonar cuando la hubo. RESULTADOS: El ajuste por edad del valor de DD supone aumentar la proporción de casos verdaderos negativos, la especificidad y el valor predictivo positivo de esta determinación. Se encuentra una correlación significativa entre el valor del DD con la extensión de la embolia pulmonar (r = 0,41, p < 0,05) pero no con la gravedad clínica del episodio. CONCLUSIONES: El ajuste del DD por la edad mejora la precisión diagnóstica de la embolia pulmonar, aunque, en nuestro entorno, su sospecha no se establece siguiendo las guías recomendadas. El valor del DD se relaciona con la extensión pero no con la gravedad de la embolia pulmonar.

In vitro and in vivo down-regulation of regulatory T cell activity with a peptide inhibitor of TGF-beta1.

Down-regulation of CD4+CD25+ regulatory T (Treg) cell function might be beneficial to enhance the immunogenicity of viral and tumor vaccines or to induce breakdown of immunotolerance. Although the mechanism of suppression used by Treg cells remains controversial, it has been postulated that TGF-beta1 mediates their immunosuppressive activity. In this study, we show that P17, a short synthetic peptide that inhibits TGF-beta1 and TGF-beta2 developed in our laboratory, is able to inhibit Treg activity in vitro and in vivo. In vitro studies demonstrate that P17 inhibits murine and human Treg-induced unresponsiveness of effector T cells to anti-CD3 stimulation, in an MLR or to a specific Ag. Moreover, administration of P17 to mice immunized with peptide vaccines containing tumor or viral Ags enhanced anti-vaccine immune responses and improved protective immunogenicity against tumor growth or viral infection or replication. When CD4+ T cells purified from OT-II transgenic mice were transferred into C57BL/6 mice bearing s.c. EG.7-OVA tumors, administration of P17 improved their proliferation, reduced the number of CD4+Foxp3+ T cells, and inhibited tumor growth. Also, P17 prevented development of immunotolerance induced by oral administration of OVA by genetically modified Lactococcus lactis in DO11.10 transgenic mice sensitized by s.c. injection of OVA. These findings demonstrate that peptide inhibitors of TGF-beta may be a valuable tool to enhance vaccination efficacy and to break tolerance against pathogens or tumor Ags.

Identification of peptide inhibitors of transforming growth factor beta 1 using a phage-displayed peptide library.

Pathologies such as liver fibrosis and scleroderma are characterized by harmful levels of transforming growth factor beta 1 (TGFbeta1). These levels could be neutralized if inhibitors of this cytokine were available. With this aim we searched for peptides with binding affinity for TGFbeta1 using a phage-displayed random 15-mer peptide library. Some peptides thus identified blocked activity of TGFbeta1 in vitro, as measured by their capacity to restore growth of Mv-1-Lu cells in presence of added TGFbeta1. Also, they inhibited TGFbeta1-dependent expression of collagen type I mRNA in liver of mice orally insulted with CCl(4). Intraperitoneal administration of 50 microg of peptide P17 (the most active 15-mer peptide, also referred to as P17(1-15)) inhibited expression of collagen type I mRNA by almost 100%. Interestingly, titration experiments showed that P17(1-12) (a peptide encompassing the first 12 amino acids of P17) was approximately four times more active than P17. These results suggest that both peptides, as well as others reported here, may be of therapeutic interest in processes requiring control of undesired high levels of TGFbeta1.