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The first transcriptional pathway regulating βIV-spectrin expression, a key angiogenic regulator of sprouting angiogenesis, is identified.

Dynamic changes in the endoplasmic reticulum (ER) morphology are central to maintaining cellular homeostasis. Microtubules (MT) facilitate the continuous remodeling of the ER network into sheets and tubules by coordinating with many ER-shaping protein complexes, although how this process is controlled by extracellular signals remains unknown. Here we report that TAK1, a kinase responsive to numerous growth factors and cytokines including TGF-β and TNF-α, triggers ER tubulation by activating αTAT1, an MT-acetylating enzyme that enhances ER-sliding. We show that this TAK1/αTAT-dependent ER remodeling promotes cell survival by actively downregulating BOK, an ER membrane-associated proapoptotic effector. While BOK is normally protected from degradation when complexed with IP3R, it is rapidly degraded upon their dissociation during the ER sheets-to-tubules conversion. These findings demonstrate a distinct mechanism of ligand-induced ER remodeling and suggest that the TAK1/αTAT pathway may be a key target in ER stress and dysfunction.

Defective angiogenesis underlies over 50 malignant, ischemic and inflammatory disorders yet long-term therapeutic applications inevitably fail, thus highlighting the need for greater understanding of the vast crosstalk and compensatory mechanisms. Based on proteomic profiling of angiogenic endothelial components, here we report βIV-spectrin, a non-erythrocytic cytoskeletal protein, as a critical regulator of sprouting angiogenesis. Early loss of endothelial-specific βIV-spectrin promotes embryonic lethality in mice due to hypervascularization and hemorrhagic defects whereas neonatal depletion yields higher vascular density and tip cell populations in developing retina. During sprouting, βIV-spectrin expresses in stalk cells to inhibit their tip cell potential by enhancing VEGFR2 turnover in a manner independent of most cell-fate determining mechanisms. Rather, βIV-spectrin recruits CaMKII to the plasma membrane to directly phosphorylate VEGFR2 at Ser984, a previously undefined phosphoregulatory site that strongly induces VEGFR2 internalization and degradation. These findings support a distinct spectrin-based mechanism of tip-stalk cell specification during vascular development.

Insulin signaling in blood vessels primarily functions to stimulate angiogenesis and maintain vascular homeostasis through the canonical PI3K and MAPK signaling pathways. However, angiogenesis is a complex process coordinated by multiple other signaling events. Here, we report a distinct crosstalk between the insulin receptor and endoglin/activin receptor-like kinase 1 (ALK1), an endothelial cell-specific TGF-β receptor complex essential for angiogenesis. While the endoglin-ALK1 complex normally binds to TGF-β or bone morphogenetic protein 9 (BMP9) to promote gene regulation via transcription factors Smad1/5, we show that insulin drives insulin receptor oligomerization with endoglin-ALK1 at the cell surface to trigger rapid Smad1/5 activation. Through quantitative proteomic analysis, we identify ependymin-related protein 1 (EPDR1) as a major Smad1/5 gene target induced by insulin but not by TGF-β or BMP9. We found endothelial EPDR1 expression is minimal at the basal state but is markedly enhanced upon prolonged insulin treatment to promote cell migration and formation of capillary tubules. Conversely, we demonstrate EPDR1 depletion strongly abrogates these angiogenic effects, indicating that EPDR1 is a crucial mediator of insulin-induced angiogenesis. Taken together, these results suggest important therapeutic implications for EPDR1 and the TGF-β pathways in pathologic angiogenesis during hyperinsulinemia and insulin resistance.

SUMMARYMitochondrial remodeling is a fundamental process underlying cellular respiration and metabolism. Here we report TAK1 as a direct regulator of mitochondrial fusion. TAK1 is activated by a variety of mitogenic factors, cytokines and environmental stimuli, which we find induces rapid fragmentation through Mfn2 inactivation. TAK1 phosphorylates Mfn2 at Ser249, which inhibits the binding of GTP required for Mfn trans-dimerization and mitochondrial membrane fusion. Accordingly, expression of Mfn2-S249 phosphomimetics (Mfn2-E/D) constitutively promote fission whereas alanine mutant (Mfn2-A) yields hyperfused mitochondria and increased bioenergetics in cells. In mice, Mfn2-E knock-in yields embryonic lethality in homozygotes whereas heterozygotes are viable but exhibit increased visceral fat accumulation despite normal body weight and cognitive/motor functions compared to wildtype and Mfn2-A mice. Mature white adipocytes isolated from mutant mice reveal cell-autonomous TAK1-related effects on mitochondrial remodeling and lipid metabolism. These results identify Mfn2-S249 as a dynamic phosphoregulatory switch of mitochondrial fusion during development and energy homeostasis.

Defective angiogenesis underlies over 50 malignant, ischemic and inflammatory disorders yet long-term therapeutic applications inevitably fail, thus highlighting the need for greater understanding of the vast crosstalk and compensatory mechanisms. Based on proteomic profiling of angiogenic endothelial components, here we report β

This study presents the chemical characteristics of flowers of Aloe vera as ingredient in food. The α-tocopherol, β-carotene, and β-sitosterol content as well as the oleoresin extraction for Aloe vera (L.) Burm f. flowers were evaluated using two agitation methods and four solvents. Then tocopherol, β-carotene, and β-sitosterol were quantified using an HPLC method. The statistical analysis revealed that the optimal extraction method for α-tocopherol was obtained by vortex agitation and acetone or ethyl acetate extraction, while for β-carotene, it was with acetone alone with no significant difference between agitation methods For β-sitosterol, the optimal method of extraction was acetone and vortex agitation. A proximate analysis of the dried Aloe flowers showed the average moisture, protein, lipid, fibre, and ash content to be 8.45%, 11.75%, 2.30%, 12.65%, and 8.07%, respectively. The best oleoresin extraction yield was by sonication using hexane as a solvent. Furthermore, Aloe flowers present an average of 30.71% inhibition for radical scavenging activity. Myristoleic acid was the most predominant fatty acid in the flowers, whereas arachidonic acid was the least abundant. Consequently, Aloe vera flowers were characterized to find their nutritional value and possible application in the food and pharmaceutical industry.

Aloin is an anthraquinone-C-glycoside present in Aloe vera. This compound is extremely variable among different species and highly depends on the growing conditions of the plants. The quantification of aloin in different extraction preparations has been a frequent problem due to the high instability of the compound. The aim of the present study is to develop and validated an analytical method for aloin detection in fresh and dry samples of Aloe barbadensis gel and latex using high performance liquid chromatography coupled to a diode array detector (HPLC-DAD). Phosphate buffered saline (pH 3) was selected as the extraction solvent. The aloin was separated using a Zorbax Eclipse AAA column (4.6 × 150 mm) at 35°C, and water and acetonitrile were used as the mobile phase at a flow rate of 0.9 mL/min. The linearity was satisfactory with a correlation coefficient greater than 0.999. Under these conditions, the method precision (relative standard deviation) was 3.71% for FL, 4.41% for dry latex, 0.81% for fresh gel and 4.42% for dry gel samples. Aloe latex was determined to have a greater amount of aloin than aloe gel. The method validation was satisfactory and exhibited adequate linearity, repeatability and accuracy. This research was financed by project PROFAPI no. 2016-0025 from Instituto Tecnológico de Sonora. info:eu-repo/semantics/publishedVersion