RESUMEN
The enzyme polyphenol oxidase (PPO) catalyzes the oxidation of phenolic compounds into highly reactive quinones. Polymerization of PPO-derived quinones causes the postharvest browning of cut or bruised fruit, but the native physiological functions of PPOs in undamaged, intact plant cells are not well understood. Walnut (Juglans regia) produces a rich array of phenolic compounds and possesses a single PPO enzyme, rendering it an ideal model to study PPO. We generated a series of PPO-silenced transgenic walnut lines that display less than 5% of wild-type PPO activity. Strikingly, the PPO-silenced plants developed spontaneous necrotic lesions on their leaves in the absence of pathogen challenge (i.e. a lesion mimic phenotype). To gain a clearer perspective on the potential functions of PPO and its possible connection to cell death, we compared the leaf transcriptomes and metabolomes of wild-type and PPO-silenced plants. Silencing of PPO caused major alterations in the metabolism of phenolic compounds and their derivatives (e.g. coumaric acid and catechin) and in the expression of phenylpropanoid pathway genes. Several observed metabolic changes point to a direct role for PPO in the metabolism of tyrosine and in the biosynthesis of the hydroxycoumarin esculetin in vivo. In addition, PPO-silenced plants displayed massive (9-fold) increases in the tyrosine-derived metabolite tyramine, whose exogenous application elicits cell death in walnut and several other plant species. Overall, these results suggest that PPO plays a novel and fundamental role in secondary metabolism and acts as an indirect regulator of cell death in walnut.
Asunto(s)
Catecol Oxidasa/metabolismo , Juglans/citología , Juglans/enzimología , Metabolismo Secundario , Muerte Celular/efectos de los fármacos , Cinamatos/metabolismo , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Silenciador del Gen/efectos de los fármacos , Juglans/efectos de los fármacos , Juglans/genética , Cinética , Metabolómica , Fenotipo , Extractos Vegetales/metabolismo , Hojas de la Planta/citología , Hojas de la Planta/efectos de los fármacos , Hojas de la Planta/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente , Propanoles/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Metabolismo Secundario/efectos de los fármacos , Metabolismo Secundario/genética , Especificidad por Sustrato/efectos de los fármacos , Tiramina/química , Tiramina/metabolismo , Tiramina/farmacologíaRESUMEN
INTRODUCTION: Hyperkalemia is a common condition, particularly in the setting of renal dysfunction. Hyperkalemia due to intentional oral potassium overdose is not commonly reported. CASE REPORT: We present a case of acute intentional potassium overdose in a patient with normal renal function resulting in significant hyperkalemia, with a maximum serum potassium concentration of 11 mEq/L. Despite an initial course complicated by various unstable cardiac rhythms, including ventricular tachycardia, ventricular fibrillation, and pulseless electrical activity, the patient was discharged from the hospital neurologically intact. Treatment for hyperkalemia included hemodialysis. DISCUSSION: The role of dialysis in potassium overdose is poorly defined. CONCLUSION: Based on this case and a review of the medical literature, we recommend hemodialysis for cases of potassium overdose with hemodynamic instability and significantly elevated serum potassium concentrations that do not respond promptly to medical therapy. Hemodialysis should also be considered in cases with underlying renal dysfunction.
Asunto(s)
Hiperpotasemia/terapia , Potasio/envenenamiento , Diálisis Renal , Intento de Suicidio , Administración Oral , Arritmias Cardíacas/etiología , Sobredosis de Droga , Femenino , Humanos , Hiperpotasemia/sangre , Hiperpotasemia/fisiopatología , Persona de Mediana Edad , Potasio/administración & dosificación , Resultado del TratamientoRESUMEN
The fully organized structure of the eukaryotic nucleosome remains unsolved, in part due to limited information regarding the binding site of the H1 or linker histone. The central globular domain of H1 is believed to interact with the nucleosome core at or near the dyad and to bind at least two strands of DNA. We utilized site-directed mutagenesis and in vivo photobleaching to identify residues that contribute to the binding of the globular domain of the somatic H1 subtype H1c to the nucleosome. As was previously observed for the H1(0) subtype, the binding residues for H1c are clustered on the surface of one face of the domain. Despite considerable structural conservation between the globular domains of these two subtypes, the locations of the binding sites identified for H1c are distinct from those of H1(0). We suggest that the globular domains of these two linker histone subtypes will bind to the nucleosome with distinct orientations that may contribute to higher order chromatin structure heterogeneity or to differences in dynamic interactions with other DNA or chromatin-binding proteins.
Asunto(s)
Histonas/metabolismo , Nucleosomas/metabolismo , Sitios de Unión , Cromatina/metabolismo , ADN/metabolismo , Regulación de la Expresión Génica , Genes Reporteros , Proteínas Fluorescentes Verdes/genética , Histonas/química , Histonas/genética , Cinética , Modelos Moleculares , Conformación Molecular , Mutagénesis , Mutagénesis Sitio-DirigidaRESUMEN
Surgeons using a fresh tissue dissection laboratory need specimens with tissue color and texture as close as possible to those of a living body. Completely unembalmed specimens kept in a cooler remain in good condition only for a few days, and then decay rapidly. Unembalmed specimens can be frozen for later use, but freezing harms their texture, and decay is suspended only for as long as they remain frozen. Since 1998, we have used a method of light embalming adapted from funeral home techniques, on over 250 cadavers used in our fresh tissue dissection laboratory. Lightly embalmed cadavers can be kept in a cooler for up to 6 weeks before use, with negligible loss of tissue quality and color. Once dissection is begun, the cadavers remain in excellent condition, free from odor, for at least two further weeks. Light embalming overcomes the practical problems seen with completely unembalmed specimens, avoids the use of freezing, and extends the range of activities that can be planned in the laboratory. This paper presents details of the light embalming technique. We assume that light embalming does not kill all transmissible pathogens.