RESUMEN
Type material of Navicula kotschyi was studied, and this species was transferred to Dorofeyukea gen. nov. as D. kotschyi comb. nov. Dorofeyukea was described on the basis of DNA sequence and morphological data. Additional species assigned to this genus that were previously included in Navicula include: D. ancisa comb. nov., D. grimmei comb. nov., D. ivatoensis comb. nov., D. orangiana comb. nov., D. rostellata comb. nov. & stat. nov., D. savannahiana comb. nov., D. tenuipunctata comb. nov., and D. texana comb. nov. All Dorofeyukea species share the same morphological features, including having a narrow stauroid fascia surrounded by 1-3 irregularly shortened striae, uniseriate, and weakly radiate striae, circular, or rectangular puncta in the striae that are covered internally by dome-shaped hymenes, presence of a pseudoseptum at each apex and absence of septa. Partial DNA sequences of SSU and rbcL loci show Dorofeuykae belongs to the clade of stauroneioid diatoms together with Stauroneis, Prestauroneis, Craticula, Karayevia, Madinithidium, Fistulifera, Parlibellus, and, possibly, Schizostauron. A new species from the monoraphid genus Madinithidium, M. vietnamica sp. nov., was described based on valve and chloroplast morphology as well as DNA sequence data.
Asunto(s)
Diatomeas , Filogenia , ARN Ribosómico 16S , Análisis de Secuencia de ADNRESUMEN
Progressive kidney diseases are often associated with scarring of the kidney's filtration unit, a condition called focal segmental glomerulosclerosis (FSGS). This scarring is due to loss of podocytes, cells critical for glomerular filtration, and leads to proteinuria and kidney failure. Inherited forms of FSGS are caused by Rac1-activating mutations, and Rac1 induces TRPC5 ion channel activity and cytoskeletal remodeling in podocytes. Whether TRPC5 activity mediates FSGS onset and progression is unknown. We identified a small molecule, AC1903, that specifically blocks TRPC5 channel activity in glomeruli of proteinuric rats. Chronic administration of AC1903 suppressed severe proteinuria and prevented podocyte loss in a transgenic rat model of FSGS. AC1903 also provided therapeutic benefit in a rat model of hypertensive proteinuric kidney disease. These data indicate that TRPC5 activity drives disease and that TRPC5 inhibitors may be valuable for the treatment of progressive kidney diseases.
Asunto(s)
Glomeruloesclerosis Focal y Segmentaria/tratamiento farmacológico , Hipertensión Renal/tratamiento farmacológico , Indazoles/farmacología , Proteinuria/tratamiento farmacológico , Canales Catiónicos TRPC/antagonistas & inhibidores , Animales , Modelos Animales de Enfermedad , Glomeruloesclerosis Focal y Segmentaria/genética , Mutación , Podocitos/efectos de los fármacos , Ratas , Ratas Endogámicas Dahl , Ratas Transgénicas , Bibliotecas de Moléculas Pequeñas , Canales Catiónicos TRPC/farmacología , Proteína de Unión al GTP rac1/genéticaRESUMEN
Tyrosine and serine/threonine signal-transduction pathways influence many aspects of cell behavior, including the spatial and temporal regulation of the actin cytoskeleton. However, little is known about how input from diverse tyrosine and serine/threonine kinases is integrated to control Rho protein crosstalk and actin remodeling, which are critically important in podocyte health and disease. Here we unveil the proteolytically-regulated, actin organizing protein synaptopodin as a coincidence detector of tyrosine versus serine/threonine phosphorylation. We show that serine/threonine and tyrosine kinases duel for synaptopodin stability versus degradation. EGFR/Src-mediated tyrosine phosphorylation of synaptopodin in podocytes promotes binding to the serine/threonine phosphatase calcineurin. This leads to the loss of 14-3-3 binding, resulting in synaptopodin degradation, Vav2 activation, enhanced Rac1 signaling, and ultimate loss of stress fibers. Our studies reveal how synaptopodin, a single proteolytically-controlled protein, integrates antagonistic tyrosine versus serine/threonine phosphorylation events for the dynamic control of the actin cytoskeleton in podocytes.
Asunto(s)
Proteínas de Microfilamentos/fisiología , Podocitos/fisiología , Serina/metabolismo , Treonina/metabolismo , Tirosina/metabolismo , Proteína de Unión al GTP rhoA/fisiología , Animales , Calcineurina/metabolismo , Células Cultivadas , Ratones , Fosforilación , Receptor Cross-Talk , Transducción de SeñalRESUMEN
Automatic systems for estimating operator fatigue have application in safety-critical environments. A system which could estimate level of fatigue from speech would have application in domains where operators engage in regular verbal communication as part of their duties. Previous studies on the prediction of fatigue from speech have been limited because of their reliance on subjective ratings and because they lack comparison to other methods for assessing fatigue. In this paper, we present an analysis of voice recordings and psychophysiological test scores collected from seven aerospace personnel during a training task in which they remained awake for 60 h. We show that voice features and test scores are affected by both the total time spent awake and the time position within each subject's circadian cycle. However, we show that time spent awake and time-of-day information are poor predictors of the test results, while voice features can give good predictions of the psychophysiological test scores and sleep latency. Mean absolute errors of prediction are possible within about 17.5% for sleep latency and 5-12% for test scores. We discuss the implications for the use of voice as a means to monitor the effects of fatigue on cognitive performance in practical applications.
RESUMEN
The ubiquitously expressed adapter proteins Nck1/2 interact with a multitude of effector molecules to regulate diverse cellular functions including cytoskeletal dynamics. Here we show that Nck1, but not Nck2, is a substrate of c-Cbl-mediated ubiquitination. We uncover lysine 178 in Nck1 as the evolutionarily conserved ubiquitin acceptor site. We previously reported that synaptopodin, a proline-rich actin-binding protein, induces stress fibres by blocking the Smurf1-mediated ubiquitination of RhoA. We now find that synaptopodin competes with c-Cbl for binding to Nck1, which prevents the ubiquitination of Nck1 by c-Cbl. Gene silencing of c-Cbl restores Nck1 protein abundance and stress fibres in synaptopodin knockdown cells. Similarly, expression of c-Cbl-resistant Nck1(K178R) or Nck2 containing the SH3 domain 2 of Nck1 restores stress fibres in synaptopodin-depleted podocytes through activation of RhoA signalling. These findings reveal proteasomal regulation as a key factor in the distinct and non-redundant effects of Nck on RhoA-mediated actin dynamics.
Asunto(s)
Actinas/metabolismo , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Proteínas Oncogénicas/metabolismo , Complejo de la Endopetidasa Proteasomal/metabolismo , Proteína de Unión al GTP rhoA/metabolismo , Proteínas Adaptadoras Transductoras de Señales/genética , Citoesqueleto/genética , Citoesqueleto/metabolismo , Activación Enzimática , Células HEK293 , Humanos , Proteínas Oncogénicas/genética , Unión Proteica , Proteolisis , Proteínas Proto-Oncogénicas c-cbl/genética , Proteínas Proto-Oncogénicas c-cbl/metabolismo , Ubiquitinación , Proteína de Unión al GTP rhoA/genéticaRESUMEN
RATIONALE: The incidence of bronchopulmonary dysplasia (BPD), a chronic lung disease of newborns, is paradoxically rising despite medical advances. We demonstrated elevated bombesin-like peptide levels in infants that later developed BPD. In the 140-day hyperoxic baboon model of BPD, anti-bombesin antibody 2A11 abrogated lung injury. OBJECTIVES: To test the hypothesis that bombesin-like peptides mediate BPD in extremely premature baboons (born at Gestational Day 125 and given oxygen pro re nata [PRN], called the 125-day PRN model), similar to "modern-day BPD." METHODS: The 125-day animals were treated with 2A11 on Postnatal Day 1 (P1), P3, and P6. On P14 and P21, lungs were inflation-fixed for histopathologic analyses of alveolarization. Regulation of angiogenesis by bombesin was evaluated using cultured pulmonary microvascular endothelial cells. MEASUREMENTS AND MAIN RESULTS: In 125-day PRN animals, urine bombesin-like peptide levels at P2-3 are directly correlated with impaired lung function at P14. Gastrin-releasing peptide (the major pulmonary bombesin-like peptide) mRNA was elevated eightfold at P1 and remained high thereafter. At P14, 2A11 reduced alveolar wall thickness and increased the percentage of secondary septa containing endothelial cells. At P21, 2A11-treated 125-day PRN animals had improved alveolarization according to mean linear intercepts and number of branch points per millimeter squared. Bombesin promoted tubulogenesis of cultured pulmonary microvascular endothelial cells, but cocultured fetal lung mesenchymal cells abrogated this effect. CONCLUSIONS: Early bombesin-like peptide overproduction in 125-day PRN animals predicted alveolarization defects weeks later. Bombesin-like peptide blockade improved septation, with the greatest effects at P21. This could have implications for preventing BPD in premature infants.