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1.
Radiologe ; 62(2): 130-139, 2022 Feb.
Artigo em Alemão | MEDLINE | ID: mdl-34997260

RESUMO

CLINICAL ISSUE: Diffuse parenchymal lung diseases include a heterogeneous group of diseases of the lung parenchyma, the alveolar spaces, the vessels and the airways, which can be triggered by various pathomechanisms, such as inflammation and fibrotic changes. Since the therapeutic approaches and prognoses differ significantly between the diseases, the correct diagnosis is of fundamental importance. In routine clinical practice, next to the patients' history, the clinical presentation, the laboratory findings and the bronchoscopy, imaging plays a central role in establishing a diagnosis. PRACTICAL RECOMMENDATIONS: The diagnosis of diffuse parenchymal lung diseases is an enormous challenge for clinicians, radiologists as well as pathologists and should therefore preferably be carried out in a multidisciplinary setting. Since patients often present with unspecific, respiratory symptoms, chest radiographs are the first imaging method used. Many patterns of diffuse parenchymal lung diseases (e.g., ground-glass opacities and consolidations), their distribution (e.g., cranial-caudal) and the presence of additional findings (e.g., mediastinal lymphadenopathy) are often already detectable on chest X­rays. However, the imaging reference standard and thus, an integral part of the assessment of diffuse parenchymal lung disease, is the chest HR-CT. In some cases, the pattern of the HR-CT is pathognomonic, in others it is unspecific for a disease, so that further diagnostic steps are necessary.


Assuntos
Doenças Pulmonares Intersticiais , Pneumopatias , Broncoscopia , Humanos , Pulmão/diagnóstico por imagem , Pneumopatias/diagnóstico por imagem , Doenças Pulmonares Intersticiais/diagnóstico por imagem , Radiografia , Tomografia Computadorizada por Raios X
2.
J Neurol ; 269(3): 1574-1590, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-34342680

RESUMO

INTRODUCTION: Adult brainstem gliomas (BSGs) are rare central nervous system tumours characterized by a highly heterogeneous clinical course. Median survival times range from 11 to 84 months. Beyond surgery, no treatment standard has been established. We investigated clinical and radiological data to assess prognostic features providing support for treatment decisions. METHODS: 34 BSG patients treated between 2000 and 2019 and aged ≥ 18 years at the time of diagnosis were retrospectively identified from the databases of the two largest Austrian Neuro-Oncology centres. Clinical data including baseline characteristics, clinical disease course, applied therapies, the outcome as well as neuroradiological and neuropathological findings were gathered and analysed. The tumour apparent diffusion coefficient (ADC), volumetry of contrast-enhancing and non-contrast-enhancing lesions were determined on magnetic resonance imaging scans performed at diagnosis. RESULTS: The median age at diagnosis was 38.5 years (range 18-71 years). Tumour progression occurred in 26/34 (76.5%) patients after a median follow up time of 19 months (range 0.9-236.2). Median overall survival (OS) and progression-free survival (PFS) was 24.1 months (range 0.9-236.2; 95% CI 18.1-30.1) and 14.5 months (range 0.7-178.5; 95% CI 5.1-23.9), respectively. Low-performance status, high body mass index (BMI) at diagnosis and WHO grading were associated with shorter PFS and OS at univariate analysis (p < 0.05, log rank test, respectively). ADC values below the median were significantly associated with shorter OS (14.9 vs 44.2 months, p = 0.018). CONCLUSION: ECOG, BMI, WHO grade and ADC values were associated with the survival prognosis of BSG patients and should be included in the prognostic assessment.


Assuntos
Neoplasias Encefálicas , Glioma , Adolescente , Adulto , Idoso , Neoplasias Encefálicas/patologia , Tronco Encefálico/patologia , Glioma/diagnóstico por imagem , Glioma/terapia , Humanos , Pessoa de Meia-Idade , Prognóstico , Estudos Retrospectivos , Centros de Atenção Terciária , Adulto Jovem
3.
Int J Mol Sci ; 22(23)2021 Dec 06.
Artigo em Inglês | MEDLINE | ID: mdl-34884966

RESUMO

Modification of kraft lignin (KL), traditionally uses harsh and energy-demanding physical and chemical processes. In this study, the potential of the bacterial laccase CotA (spore coating protein A) for oxidation of KL under mild conditions was assessed. Thereby, the efficiency of CotA to oxidize both softwood and hardwood KL of varying purity at alkaline conditions was examined. For the respective type of wood, the highest oxidation activity by CotA was determined for the medium ash content softwood KL (MA_S) and the medium ash content hardwood KL (MA_H), respectively. By an up to 95% decrease in fluorescence and up to 65% in phenol content coupling of the structural lignin units was indicated. These results correlated with an increase in viscosity and molecular weight, which increased nearly 2 and 20-fold for MA_H and about 1.3 and 6.0-fold for MA_S, respectively. Thus, this study confirms that the CotA laccase can oxidize a variety of KL at alkaline conditions, while the origin and purity of KL were found to have a major impact on the efficiency of oxidation. Under the herein tested conditions, it was observed that the MA_H KL showed the highest susceptibility to CotA oxidation when compared to the other hardwood KLs and the softwood KLs. Therefore, this could be a viable method to produce sustainable resins and adhesives.


Assuntos
Bactérias/enzimologia , Proteínas de Bactérias/metabolismo , Lacase/metabolismo , Lignina/química , Proteínas de Bactérias/genética , Lacase/genética , Peso Molecular , Oxirredução
4.
Ther Adv Med Oncol ; 13: 17588359211009002, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33995593

RESUMO

BACKGROUND: Dual human epidermal growth factor receptor 2 (HER2) blockade with trastuzumab and pertuzumab (TP) is a standard therapy of metastatic and localized HER2-positive breast cancer (BC), but its activity in breast cancer brain metastases (BCBM) is unknown. METHODS: Patients with HER2-positive BCBM were identified from the Vienna Brain Metastasis Registry and clinical data including patient characteristics, therapies and overall survival (OS) were obtained. Patients were grouped into 'TP', 'other-HER2-targeted therapy' and 'no-HER2-targeted therapy' according to received first-line systemic therapy after diagnosis of BCBM. Radiological re-assessment of intracranial lesions was performed in patients treated with TP as systemic first-line therapy according to RANO response criteria for brain metastases (BM). RESULTS: A total of 252 HER2-positive BC patients with BM were available for this analysis. Patients treated with TP as systemic first-line therapy after diagnosis of BM had a significantly longer OS compared with treatment with other-HER2-targeted therapy and no-HER2-targeted therapy (44 versus 17 versus 3 months, p < 0.001; log-rank test). Among radiologically re-assessed patients treated with TP as systemic first-line therapy after diagnosis of BM, 5/14 patients (35.7%) had complete intracranial remission (CR), 8/14 patients (57.1%) partial intracranial remission (PR), 1/14 patients (7.1%) stable intracranial disease (SD) and 0/14 patients (0.0%) progressive intracranial disease (PD) as best response resulting in an intracranial objective response rate (iORR) of 92.9% and an intracranial clinical benefit rate (iCBR) of 100.0%. CONCLUSION: First-line therapy with dual HER2-inhibition of TP after BM diagnosis was associated with the longest median OS times in patients with BCBM.

6.
World Neurosurg ; 143: e365-e373, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32730972

RESUMO

BACKGROUND: Extent of resection (EOR) constitutes a crucial factor for patient prognosis in surgery of brain metastases (BMs). According to early studies using postoperative magnetic resonance imaging (MRI), an unexpected residual tumor was not uncommon. Knowledge of potential risk factors for incomplete BM resection would be of major importance to optimize surgical strategies. The aim of this study was to evaluate EOR in a large cohort and analyze potential risk factors for incomplete BM resection. METHODS: Patients with BM resection and available postoperative MRI were included. Intraoperative estimation of EOR by the neurosurgeon was noted. Additionally, EOR was determined by postoperative MRI. Potential risk factors for incomplete resection were investigated. RESULTS: There were 145 patients with 163 BMs included. According to postoperative MRI, complete resection was achieved in 103 (63%) BMs, and resection was incomplete in 44 (27%) BMs. Postoperative MRI detected unexpected residual tumor in 32 (25%) BMs, and a misjudgment of the EOR by the neurosurgeon was found in 29% of cases. Regarding risk factors for incomplete resection, preoperative tumor volume was significantly larger in incompletely resected BMs compared with completely resected BMs (P = 0.011). All other analyzed risk factors had no significant influence on EOR. CONCLUSIONS: Our data indicate that postoperative MRI is able to detect a high portion of unexpected residual tumors after surgery of BMs. Preoperative tumor volume in particular represents an important risk factor for incomplete resection, and hence neurosurgeons should pay special attention to avoid residual tumor tissue.


Assuntos
Neoplasias Encefálicas/diagnóstico por imagem , Neoplasias Encefálicas/cirurgia , Imageamento por Ressonância Magnética/métodos , Neoplasia Residual/diagnóstico por imagem , Neoplasia Residual/cirurgia , Cuidados Pós-Operatórios/métodos , Adulto , Idoso , Estudos de Coortes , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Fatores de Risco
7.
J Clin Med ; 9(5)2020 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-32354003

RESUMO

Temporal muscle thickness (TMT) was investigated as a novel surrogate marker on MRI examinations of the brain, to detect patients who may be at risk for sarcopenia. TMT was analyzed in a retrospective, normal collective cohort (n = 624), to establish standard reference values. These reference values were correlated with grip strength measurements and body mass index (BMI) in 422 healthy volunteers and validated in a prospective cohort (n = 130) of patients with various neurological disorders. Pearson correlation revealed a strong association between TMT and grip strength (retrospective cohort, ρ = 0.746; p < 0.001; prospective cohort, ρ = 0.649; p < 0.001). A low or no association was found between TMT and age (retrospective cohort, R2 correlation coefficient 0.20; p < 0.001; prospective cohort, ρ = -0.199; p = 0.023), or BMI (retrospective cohort, ρ = 0.116; p = 0.042; prospective cohort, ρ = 0.227; p = 0.009), respectively. Male patients with temporal wasting and unintended weight loss, respectively, showed significantly lower TMT values (p = 0.04 and p = 0.015, unpaired t-test). TMT showed a high correlation with muscle strength in healthy individuals and in patients with various neurological disorders. Therefore, TMT should be integrated into the diagnostic workup of neurological patients, to prevent, delay, or treat sarcopenia.

8.
Nat Commun ; 10(1): 5516, 2019 12 04.
Artigo em Inglês | MEDLINE | ID: mdl-31797871

RESUMO

Arabidopsis PIN2 protein directs transport of the phytohormone auxin from the root tip into the root elongation zone. Variation in hormone transport, which depends on a delicate interplay between PIN2 sorting to and from polar plasma membrane domains, determines root growth. By employing a constitutively degraded version of PIN2, we identify brassinolides as antagonists of PIN2 endocytosis. This response does not require de novo protein synthesis, but involves early events in canonical brassinolide signaling. Brassinolide-controlled adjustments in PIN2 sorting and intracellular distribution governs formation of a lateral PIN2 gradient in gravistimulated roots, coinciding with adjustments in auxin signaling and directional root growth. Strikingly, simulations indicate that PIN2 gradient formation is no prerequisite for root bending but rather dampens asymmetric auxin flow and signaling. Crosstalk between brassinolide signaling and endocytic PIN2 sorting, thus, appears essential for determining the rate of gravity-induced root curvature via attenuation of differential cell elongation.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Brassinosteroides/metabolismo , Gravitropismo/fisiologia , Raízes de Plantas/metabolismo , Arabidopsis/efeitos dos fármacos , Transporte Biológico/efeitos dos fármacos , Brassinosteroides/farmacologia , Endocitose/efeitos dos fármacos , Gravitropismo/efeitos dos fármacos , Ácidos Indolacéticos/metabolismo , Meristema/efeitos dos fármacos , Meristema/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Reguladores de Crescimento de Plantas/farmacologia , Raízes de Plantas/efeitos dos fármacos , Transdução de Sinais , Esteroides Heterocíclicos/metabolismo , Esteroides Heterocíclicos/farmacologia
9.
PLoS One ; 13(11): e0207849, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30496307

RESUMO

OBJECTIVES: This study aimed to assess the correlation of temporal muscle thickness (TMT), measured on routine cranial magnetic resonance (MR) images, with lumbar skeletal muscles obtained on computed tomography (CT) images in brain metastasis patients to establish a new parameter estimating skeletal muscle mass on brain MR images. METHODS: We retrospectively analyzed the cross-sectional area (CSA) of skeletal muscles at the level of the third lumbar vertebra on computed tomography scans and correlated these values with TMT on MR images of the brain in two independent cohorts of 93 lung cancer and 61 melanoma patients (overall: 154 patients) with brain metastases. RESULTS: Pearson correlation revealed a strong association between mean TMT and CSA in lung cancer and melanoma patients with brain metastases (0.733; p<0.001). The two study cohorts did not differ significantly in patient characteristics, including age (p = 0.661), weight (p = 0.787), and height (p = 0.123). However, TMT and CSA measures differed significantly between male and female patients in both lung cancer and melanoma patients with brain metastases (p<0.001). CONCLUSION: Our data indicate that TMT, measured on routine cranial MR images, is a useful surrogate parameter for the estimation of skeletal muscle mass in patients with brain metastases. Thus, TMT may be useful for prognostic assessment, treatment considerations, and stratification or a selection factor for clinical trials in patients with brain metastases. Further studies are needed to assess the association between TMT and clinical frailty parameters, and the usefulness of TMT in patients with primary brain tumors.


Assuntos
Neoplasias Encefálicas/patologia , Neoplasias Encefálicas/secundário , Vértebras Lombares , Músculo Esquelético/patologia , Neoplasias Encefálicas/diagnóstico por imagem , Estudos Transversais , Feminino , Humanos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Músculo Esquelético/diagnóstico por imagem , Estudos Retrospectivos , Tomografia Computadorizada por Raios X
10.
J Neurooncol ; 140(1): 173-178, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30008154

RESUMO

OBJECTIVES: The purpose of this study was to evaluate the prognostic relevance of temporal muscle thickness (TMT) in melanoma patients with newly diagnosed brain metastases. METHODS: TMT was retrospectively assessed in 146 melanoma patients with newly diagnosed brain metastases on cranial magnetic resonance images. Chart review was used to retrieve clinical parameters, including disease-specific graded prognostic assessment (DS-GPA) and survival times. RESULTS: Patients with a TMT > median showed a statistically significant increase in survival time (13 months) compared to patients with a TMT < median (5 months; p < 0.001; log rank test). A Cox regression model revealed that the risk of death was increased by 27.9% with every millimeter reduction in TMT. In the multivariate analysis, TMT (HR 0.724; 95% 0.642-0.816; < 0.001) and DS-GPA (HR 1.214; 95% CI 1.023-1.439; p = 0.026) showed a statistically significant correlation with overall survival. CONCLUSION: TMT is an independent predictor of survival in melanoma patients with brain metastases. This parameter may aid in patient selection for clinical trials or to the choice of different treatment options based on the determination of frail patient populations.


Assuntos
Neoplasias Encefálicas/patologia , Neoplasias Encefálicas/secundário , Melanoma/patologia , Músculo Temporal/patologia , Adolescente , Adulto , Neoplasias Encefálicas/diagnóstico por imagem , Feminino , Humanos , Imageamento por Ressonância Magnética , Masculino , Melanoma/diagnóstico por imagem , Prognóstico , Análise de Regressão , Estudos Retrospectivos , Análise de Sobrevida , Tomógrafos Computadorizados , Adulto Jovem
11.
Exp Physiol ; 102(6): 650-662, 2017 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-28370799

RESUMO

NEW FINDINGS: What is the central question of this study? Knockdown of UCP2 reduces mitochondrial Ca2+ uptake. This suggests that Ucp2 knockout mice need to have additional effects on cytosolic Ca2+ handling to prevent Ca2+ overload. However, the specific mechanisms and their impact on cardiac electrophysiology remain speculative. What is the main finding and its importance? In Ucp2 knockout mice, decreased mitochondrial Ca2+ uptake is compensated for by functional inhibition of L-type Ca2+ channels and resultant shortening of action potential duration. UCP2-dependent modulations have a major impact on cardiac electrophysiology, resulting in alterations of ECG characteristics and a higher susceptibility to Ca2+ -mediated ventricular arrhythmias. Uncoupling protein 2 (mitochondrial, proton carrier) (UCP2) belongs to a superfamily of mitochondrial ion transporters. Owing to its beneficial influence on production of reactive oxygen species, it is suggested to reduce cardiac ischaemia-reperfusion injury. Recent studies have uncovered its ability to regulate mitochondrial Ca2+ uptake and therefore to influence cardiac cytosolic Ca2+ handling, indicating compensatory pathways to avoid toxic Ca2+ overload in Ucp2 knockout (Ucp2-/- ) mice. However, the specific mechanisms and their impact on cardiac electrophysiology remain speculative. Molecular analyses, whole-cell patch clamp in cardiomyocytes and ECG studies were performed in Ucp2-/- and wild-type (WT) control mice. Furthermore, to explore the impact on cardiac arrhythmogenicity, ECG monitoring was performed in basal conditions and during Ca2+ -mediated stress using Bay K 8644. Although cardiac ryanodine receptor 2, NCX1, L-type Ca2+ channel (LTCC) and SERCA2a expression were not altered, Ucp2-/- mice revealed major variations in cardiac electrophysiology. The LTCC current and APD90 were decreased in Ucp2-/- mice, indicating compensatory mechanisms. Furthermore, in Ucp2-/- mice, an increased slope factor of action potential upstrokes and more hyperpolarized resting membrane potential were measured, suggesting variations in cardiac excitability. In agreement with alterations of cellular physiology in Ucp2-/- mice, reductions in PR and QRS as well as shortening of the QTc interval were noted in ECG recordings. Importantly, an increased incidence of cellular after-depolarizations and more pronounced susceptibility to Ca2+ -mediated arrhythmias were observed. Furthermore, although expression of UCP3 was not different, levels of PRMT1 were significantly higher in Ucp2-/- mice. Our observations indicate compensatory mechanisms by which Ucp2-/- mice prevent toxic cytosolic Ca2+ overload. UCP2-dependent modulations have a major impact on cardiac electrophysiology and influence susceptibility to Ca2+ -mediated ventricular arrhythmias.


Assuntos
Arritmias Cardíacas/metabolismo , Cálcio/metabolismo , Proteína Desacopladora 2/metabolismo , Animais , Citosol/metabolismo , Eletrofisiologia/métodos , Masculino , Potencial da Membrana Mitocondrial/fisiologia , Mitocôndrias Cardíacas/metabolismo , Proteínas Mitocondriais/metabolismo , Miócitos Cardíacos/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Traumatismo por Reperfusão/metabolismo , Canal de Liberação de Cálcio do Receptor de Rianodina/metabolismo , ATPases Transportadoras de Cálcio do Retículo Sarcoplasmático/metabolismo , Trocador de Sódio e Cálcio/metabolismo
12.
Sci Rep ; 6: 32196, 2016 08 24.
Artigo em Inglês | MEDLINE | ID: mdl-27553690

RESUMO

Directional transport of auxin is essential for plant development, with PIN auxin transport proteins representing an integral part of the machinery that controls hormone distribution. However, unlike the rapidly emerging framework of molecular determinants regulating PIN protein abundance and subcellular localization, insights into mechanisms controlling PIN transcription are still limited. Here we describe PIN2 PROMOTER BINDING PROTEIN 1 (PPP1), an evolutionary conserved plant-specific DNA binding protein that acts on transcription of PIN genes. Consistent with PPP1 DNA-binding activity, PPP1 reporter proteins are nuclear localized and analysis of PPP1 null alleles and knockdown lines indicated a function as a positive regulator of PIN expression. Furthermore, we show that ppp1 pleiotropic mutant phenotypes are partially reverted by PIN overexpression, and results are presented that underline a role of PPP1-PIN promoter interaction in PIN expression control. Collectively, our findings identify an elementary, thus far unknown, plant-specific DNA-binding protein required for post-embryonic plant development, in general, and correct expression of PIN genes, in particular.


Assuntos
Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/genética , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo , Proteínas de Arabidopsis/química , Sítios de Ligação , Núcleo Celular/metabolismo , Simulação por Computador , Citoplasma/metabolismo , Proteínas de Ligação a DNA/química , Regulação da Expressão Gênica de Plantas , Meristema/fisiologia , Filogenia , Plantas Geneticamente Modificadas , Regiões Promotoras Genéticas , Domínios Proteicos , Proteínas de Ligação a RNA/química
13.
PLoS One ; 11(2): e0148359, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26849136

RESUMO

INTRODUCTION: The possible role of UCP2 in modulating mitochondrial Ca2+-uptake (mCa2+-uptake) via the mitochondrial calcium uniporter (MCU) is highly controversial. METHODS: Thus, we analyzed mCa2+-uptake in isolated cardiac mitochondria, MCU single-channel activity in cardiac mitoplasts, dual Ca2+-transients from mitochondrial ((Ca2+)m) and intracellular compartment ((Ca2+)c) in the whole-cell configuration in cardiomyocytes of wild-type (WT) and UCP2-/- mice. RESULTS: Isolated mitochondria showed a Ru360 sensitive mCa2+-uptake, which was significantly decreased in UCP2-/- (229.4±30.8 FU vs. 146.3±23.4 FU, P<0.05). Single-channel registrations confirmed a Ru360 sensitive voltage-gated Ca2+-channel in mitoplasts, i.e. mCa1, showing a reduced single-channel activity in UCP2-/- (Po,total: 0.34±0.05% vs. 0.07±0.01%, P<0.05). In UCP2-/- cardiomyocytes (Ca2+)m was decreased (0.050±0.009 FU vs. 0.021±0.005 FU, P<0.05) while (Ca2+)c was unchanged (0.032±0.002 FU vs. 0.028±0.004 FU, P>0.05) and transsarcolemmal Ca2+-influx was inhibited suggesting a possible compensatory mechanism. Additionally, we observed an inhibitory effect of ATP on mCa2+-uptake in WT mitoplasts and (Ca2+)m of cardiomyocytes leading to an increase of (Ca2+)c while no ATP dependent effect was observed in UCP2-/-. CONCLUSION: Our results indicate regulatory effects of UCP2 on mCa2+-uptake. Furthermore, we propose, that previously described inhibitory effects on MCU by ATP may be mediated via UCP2 resulting in changes of excitation contraction coupling.


Assuntos
Cálcio/metabolismo , Canais Iônicos/metabolismo , Mitocôndrias Cardíacas/metabolismo , Proteínas Mitocondriais/metabolismo , Trifosfato de Adenosina/metabolismo , Trifosfato de Adenosina/farmacologia , Animais , Canais de Cálcio/metabolismo , Citosol/efeitos dos fármacos , Citosol/metabolismo , Relação Dose-Resposta a Droga , Canais Iônicos/genética , Masculino , Potencial da Membrana Mitocondrial , Camundongos Mutantes , Mitocôndrias Cardíacas/efeitos dos fármacos , Proteínas Mitocondriais/genética , Miócitos Cardíacos/metabolismo , Técnicas de Patch-Clamp , Compostos de Rutênio/administração & dosagem , Compostos de Rutênio/farmacologia , Sarcolema/metabolismo , Proteína Desacopladora 2
14.
Cell Rep ; 11(4): 516-26, 2015 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-25892242

RESUMO

Polar transport of the phytohormone auxin throughout plants shapes morphogenesis and is subject to stringent and specific control. Here, we identify basic cellular activities connected to translational control of gene expression as sufficient to specify auxin-mediated development. Mutants in subunits of Arabidopsis Elongator, a protein complex modulating translational efficiency via maturation of tRNAs, exhibit defects in auxin-controlled developmental processes, associated with reduced abundance of PIN-formed (PIN) auxin transport proteins. Similar anomalies are observed upon interference with tRNA splicing by downregulation of RNA ligase (AtRNL), pointing to a general role of tRNA maturation in auxin signaling. Elongator Protein 6 (ELP6) and AtRNL expression patterns underline an involvement in adjusting PIN protein levels, whereas rescue of mutant defects by auxin indicates rate-limiting activities in auxin-controlled organogenesis. This emphasizes mechanisms in which auxin serves as a bottleneck for plant morphogenesis, translating common cellular activities into defined developmental readouts.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Ácidos Indolacéticos/metabolismo , Processamento Pós-Transcricional do RNA , RNA de Transferência/metabolismo , Proteínas de Ligação a RNA/metabolismo , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/genética , Proteínas de Membrana Transportadoras/genética , Proteínas de Membrana Transportadoras/metabolismo , Proteínas de Ligação a RNA/genética
15.
Methods Mol Biol ; 1209: 233-49, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25117288

RESUMO

Reversible, covalent modification by the small protein ubiquitin acts in a variety of pathways controlling protein fate in virtually all aspects of cellular function. For example, ubiquitylation of plasma membrane proteins modulates their intracellular sorting and turnover, thereby decisively influencing crosstalk between cells and their environment. In recent years, experimental work performed with the model plant Arabidopsis thaliana demonstrated ubiquitylation of a number of plasma membrane proteins, including the auxin efflux carrier protein PIN2. By using solubilized membrane protein immunoprecipitation assays, we established quantitative approaches, suitable for analysis of PIN2 ubiquitylation and variations therein. Applicability of this robust approach is not restricted to PIN auxin carriers, but could be extended to analysis of further plant membrane proteins that are controlled by variations in their ubiquitylation status.


Assuntos
Proteínas de Arabidopsis/metabolismo , Biologia Molecular/métodos , Transporte Proteico , Ubiquitinação/genética , Proteínas de Arabidopsis/isolamento & purificação , Imunoprecipitação , Ácidos Indolacéticos/metabolismo
16.
Life (Basel) ; 3(4): 538-49, 2013 Dec 17.
Artigo em Inglês | MEDLINE | ID: mdl-25369885

RESUMO

The problem of how life began can be considered as a matter of basic chemistry. How did the molecules of life arise from non-biological chemistry? Stanley Miller's famous experiment in 1953, in which he produced amino acids under simulated early Earth conditions, was a huge leap forward in our understanding of this problem. Our research first simulated early Earth conditions based on Miller's experiment and we then repeated the experiment using Titan post-impact conditions. We simulated conditions that could have existed on Titan after an asteroid strike. Specifically, we simulated conditions after a potential strike in the subpolar regions of Titan that exhibit vast methane-ethane lakes. If the asteroid or comet was of sufficient size, it would also puncture the icy crust and bring up some of the subsurface liquid ammonia-water mixture. Since, O'Brian, Lorenz and Lunine showed that a liquid water-ammonia body could exist between about 102-104 years on Titan after an asteroid impact we modified our experimental conditions to include an ammonia-water mixture in the reaction medium. Here we report on the resulting amino acids found using the Titan post-impact conditions in a classical Miller experimental reaction set-up and how they differ from the simulated early Earth conditions.

17.
Plant Signal Behav ; 7(10): 1271-3, 2012 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-22902683

RESUMO

Plant tropisms are decisively influenced by dynamic adjustments in spatiotemporal distribution of the growth regulators auxin. Polar auxin transport requires activity of PIN-type auxin carrier proteins, with their distribution at the plasma membrane significantly contributing to the directionality of auxin flow. Control of PIN protein distribution involves regulation of their endocytosis and further sorting into the lytic vacuole for degradation and recently, protein ubiquitylation has been demonstrated to control degradative sorting of plasma membrane proteins in plants. ( 1) (-) ( 6) Here we show dynamic adjustments in PIN2 ubiquitylation in gravity-stimulated roots, a response that coincides with establishment of a lateral PIN2 expression gradient. Our results imply that perception and transduction of gravity signals triggers differential ubiquitylation of PIN2, which might feed back on the coordination of auxin distribution in root meristems.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/fisiologia , Gravitação , Ácidos Indolacéticos/metabolismo , Raízes de Plantas/fisiologia , Ubiquitinação , Western Blotting , Imunoprecipitação
18.
Proc Natl Acad Sci U S A ; 109(21): 8322-7, 2012 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-22556266

RESUMO

Cross-talk between plant cells and their surroundings requires tight regulation of information exchange at the plasma membrane (PM), which involves dynamic adjustments of PM protein localization and turnover to modulate signal perception and solute transport at the interface between cells and their surroundings. In animals and fungi, turnover of PM proteins is controlled by reversible ubiquitylation, which signals endocytosis and delivery to the cell's lytic compartment, and there is emerging evidence for related mechanisms in plants. Here, we describe the fate of Arabidopsis PIN2 protein, required for directional cellular efflux of the phytohormone auxin, and identify cis- and trans-acting mediators of PIN2 ubiquitylation. We demonstrate that ubiquitin acts as a principal signal for PM protein endocytosis in plants and reveal dynamic adjustments in PIN2 ubiquitylation coinciding with variations in vacuolar targeting and proteolytic turnover. We show that control of PIN2 proteolytic turnover via its ubiquitylation status is of significant importance for auxin distribution in root meristems and for environmentally controlled adaptations of root growth. Moreover, we provide experimental evidence indicating that PIN2 vacuolar sorting depends on modification specifically by lysine(63)-linked ubiquitin chains. Collectively, our results establish lysine(63)-linked PM cargo ubiquitylation as a regulator of polar auxin transport and adaptive growth responses in higher plants.


Assuntos
Adaptação Fisiológica/fisiologia , Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Ácidos Indolacéticos/metabolismo , Raízes de Plantas/metabolismo , Ubiquitinação/fisiologia , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/genética , Endocitose/fisiologia , Genótipo , Gravitropismo/fisiologia , Lisina/metabolismo , Raízes de Plantas/crescimento & desenvolvimento , Ubiquitina-Proteína Ligases/genética , Ubiquitina-Proteína Ligases/metabolismo , Vacúolos/metabolismo
19.
Dev Cell ; 22(3): 678-85, 2012 Mar 13.
Artigo em Inglês | MEDLINE | ID: mdl-22421050

RESUMO

Growth and development are coordinated by an array of intercellular communications. Known plant signaling molecules include phytohormones and hormone peptides. Although both classes can be implicated in the same developmental processes, little is known about the interplay between phytohormone action and peptide signaling within the cellular microenvironment. We show that genes coding for small secretory peptides, designated GOLVEN (GLV), modulate the distribution of the phytohormone auxin. The deregulation of the GLV function impairs the formation of auxin gradients and alters the reorientation of shoots and roots after a gravity stimulus. Specifically, the GLV signal modulates the trafficking dynamics of the auxin efflux carrier PIN-FORMED2 involved in root tropic responses and meristem organization. Our work links the local action of secretory peptides with phytohormone transport.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Proteínas de Transporte/metabolismo , Gravitropismo , Ácidos Indolacéticos/metabolismo , Arabidopsis/genética , Proteínas de Transporte/genética , Microambiente Celular/fisiologia , Regulação da Expressão Gênica de Plantas , Raízes de Plantas/genética , Raízes de Plantas/metabolismo , Raízes de Plantas/fisiologia , Transdução de Sinais/genética
20.
Orig Life Evol Biosph ; 41(6): 545-52, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22139520

RESUMO

A radiative convective model to calculate the width and the location of the life supporting zone (LSZ) for different, alternative solvents (i.e. other than water) is presented. This model can be applied to the atmospheres of the terrestrial planets in the solar system as well as (hypothetical, Earth-like) terrestrial exoplanets. Cloud droplet formation and growth are investigated using a cloud parcel model. Clouds can be incorporated into the radiative transfer calculations. Test runs for Earth, Mars and Titan show a good agreement of model results with observations.


Assuntos
Atmosfera , Meio Ambiente Extraterreno/química , Modelos Químicos , Origem da Vida , Planetas , Evolução Planetária , Exobiologia
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