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1.
J Plast Reconstr Aesthet Surg ; 95: 124-126, 2024 Jun 11.
Artigo em Inglês | MEDLINE | ID: mdl-38905788

RESUMO

Osteotomy is often necessary for the repair of post-traumatic nasal bone deformities. Typically, tools such as chisels are used for osteotomies; however, we performed osteotomies using Kirschner wires without making a skin incision. Between April 2011 and July 2022, we performed rhinoplasty with external perforated osteotomy using Kirschner wires in 13 patients with post-traumatic nasal bone deformities (9 males and 4 females; mean age, 34 years, range 12-51 years), all of whom exhibited improvement, with only one case showing mild residual cosmetic deformity. None of the patients requested further revision, and all were satisfied with their functional results. The non-incisional external perforated technique is a reasonable method that allows for bone osteotomies along the fracture line and is well-controlled, predictable, and reproducible.

2.
Yakugaku Zasshi ; 144(7): 741-748, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38945848

RESUMO

In 2020, the Japan Community Healthcare Organization (JCHO) Hoshigaoka Medical Center started providing information to community pharmacies about patients admitted to the acute care ward using discharge medication summaries (the summaries). We conducted an online self-recording survey of 149 pharmacies belonging to the Hirakata City Pharmacists Association to clarify the usability of the summaries, any related issues, and to further discuss future collaboration between hospitals and pharmacies. 46 pharmacies have received the summaries in the past, of which 44 pharmacies answered that they have utilized the summaries with patient instruction and prescription queries of doctors. However, two pharmacies responded they did not utilize the summaries, and the reasons were (a) the information was not timely and (b) patients whom the discharge medical summary was sent for did not come to the pharmacy. There were some requests regarding the summaries such as, "I would like to know what kind of information hospital pharmacists want from community pharmacists." Preference for sharing information other than the summaries (e.g., online tools) with hospital pharmacists was related to whether the pharmacy was providing home pharmaceutical visit services. The survey revealed that, in addition to the usability of the summaries, there are also events that prevent them from being utilized. Some of the challenges include the timing of sending the summaries, the accurate identification of the family pharmacy and the communication of follow-up after discharge from hospital. Collaborating with pharmacies providing home pharmaceutical visit services would be beneficial in creating new system of bidirectional information sharing.


Assuntos
Serviços Comunitários de Farmácia , Alta do Paciente , Farmacêuticos , Serviço de Farmácia Hospitalar , Humanos , Japão , Inquéritos e Questionários , Sumários de Alta do Paciente Hospitalar
3.
Intern Med ; 2023 Dec 18.
Artigo em Inglês | MEDLINE | ID: mdl-38104988

RESUMO

We herein report a 61-year-old woman with a history of mitral valve replacement for rheumatic fever who presented with crural edema and ascites. Computed tomography showed massive left atrial (LA) calcification involving the interatrial septum, termed "coconut atrium." Catheterization revealed not only pulmonary artery hypertension but also a large V-wave in the pulmonary artery wedge pressure waveform and a dip-and-plateau pattern of right ventricular pressure. Three-dimensional transthoracic echocardiography confirmed the early attainment of peak LA volume and a decreased LA expansion index. Stiff LA syndrome due to coconut LA results in the development of restrictive right ventricular physiology.

4.
ESC Heart Fail ; 10(3): 2031-2041, 2023 06.
Artigo em Inglês | MEDLINE | ID: mdl-37057311

RESUMO

AIMS: In patients with aortic stenosis (AS), the coronary flow reserve decreases even in the absence of epicardial coronary artery stenosis. Systolic coronary flow reversal (SFR) reflecting reduced coronary microcirculation, often seen in patients with severe AS, has a potential negative impact on the pathogenesis of cardiac dysfunction. However, there are limited data on the relationship between the severity of AS and SFR, as well as on the benefits of transcatheter aortic valve implantation (TAVI). The aim of this study was to evaluate the relationship between the severity of AS and efficacy of TAVI in improving SFR. METHODS AND RESULTS: Consecutive patients with AS who had undergone TAVI using transoesophageal echocardiography (TEE) from November 2020 to February 2022 were prospectively enrolled. Coronary flow in the left anterior descending artery as well as the aortic valve peak velocities, and the mean aortic valve pressure gradients (AVPGs), indicating the severity of AS, were measured using intraprocedural TEE before and after TAVI. The following parameters were measured as coronary flow: systolic and diastolic peak velocity (cm/s) and systolic and diastolic velocity-time integral (VTI) (cm). SFR was defined as the presence of a reversal coronary flow component in systole. The enrolled patients were classified into two groups according to the presence or absence of SFR before TAVI. A total of 25 patients were included: 13 had SFR and 12 who had no SFR, before TAVI. Patients with SFR had significantly higher aortic valve peak velocities (451.1 ± 45.9 vs. 372.1 ± 52.1 cm/s; P < 0.001) and mean AVPGs (49.2 ± 14.5 vs. 30.3 ± 11.6 mmHg; P = 0.002) than those without. The optimal binary cut-off aortic valve peak velocity values and the mean AVPG associated with the presence of SFR before TAVI were >410.0 cm/s (specificity, 75.0%; sensitivity, 92.3%) and >37.4 mmHg (specificity, 83.3%; sensitivity, 92.3%), respectively. After TAVI, SFR immediately disappeared in 11 of 13 patients with SFR (84.6%). Overall, the systolic coronary VTI significantly increased after TAVI (2.0 ± 4.7 vs. 6.4 ± 3.2 cm, P < 0.001), and this increase was greater in patients with SFR than in those without SFR before TAVI (interaction P = 0.035). CONCLUSIONS: SFR was found to be associated with the severity of AS and with a greater increase in systolic coronary flow immediately after TAVI.


Assuntos
Estenose da Valva Aórtica , Substituição da Valva Aórtica Transcateter , Humanos , Substituição da Valva Aórtica Transcateter/efeitos adversos , Circulação Coronária , Cateterismo Cardíaco , Estenose da Valva Aórtica/diagnóstico , Estenose da Valva Aórtica/cirurgia , Estenose da Valva Aórtica/complicações , Valva Aórtica/diagnóstico por imagem , Valva Aórtica/cirurgia
5.
Cureus ; 15(12): e50081, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-38186451

RESUMO

Group A streptococcus pyogenes (GAS) is a common organism that can cause upper respiratory infections. We encountered a case where GAS caused infective endocarditis (IE) due to an entry from the vagina. In this case, although echocardiography was negative, we were able to make a diagnosis of IE based on the 2023 Duke International Society for Cardiovascular Infectious Diseases Criteria, and we started antimicrobial therapy for IE. However, the patient subsequently developed persistent abdominal pain, which was atypical; hence, we reviewed the differential diagnosis. It is difficult to locate the primary site of infection because GAS rarely causes vaginal infections, and vaginal infections rarely cause IE. This case highlights the significance of revisiting medical history and the value of using a system 3 approach to refine diagnostic directions.

6.
JBJS Case Connect ; 12(4)2022 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-36563172

RESUMO

CASE: A 70-year-old woman presented with severe trunk deformity, gait disturbance, and bilateral hip pain. Radiography and magnetic resonance imaging revealed severe spinal kyphosis, bilateral osteoarthritis of the hip joints, and lumbar canal stenosis. A 2-stage corrective surgery for the treatment of spinal kyphosis was performed. Subsequently, the spinopelvic alignment changed remarkably. The spinal symptoms and radiographic parameters improved postoperatively, and bilateral hip pain and range of motion also showed improvement. CONCLUSION: Corrective surgery for the treatment of degenerative spinal kyphosis affected the spinopelvic alignment, and the symptoms of simultaneous coxarthrosis also improved after the corrective surgery.


Assuntos
Cifose , Osteoartrite do Quadril , Feminino , Humanos , Idoso , Osteoartrite do Quadril/cirurgia , Coluna Vertebral/cirurgia , Pelve , Cifose/cirurgia , Dor
7.
Clin Case Rep ; 10(8): e6251, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35999981

RESUMO

Emphysematous pyelonephritis is a severe necrotizing infection that occurs predominately among individuals with diabetes mellitus. The mortality rate is >10% only medical therapy. We report a case of emphysematous pyelonephritis caused by Klebsiella pneumoniae, precipitated by faulty catheter insertion. The patient recovered after the replacement of catheter and administration of antibiotics.

8.
Clin Case Rep ; 10(4): e05673, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35414912

RESUMO

Kikuchi disease is common among Asians and women aged about 30 years. General symptoms include cervical lymphadenopathy and fever; 64% of the cases are self-limiting. However, this case is axillary lymphadenopathy, which is the main symptom in males. Thus, Kikuchi disease may be considered in the differential diagnosis of axillary lymphadenopathy.

9.
J Nat Med ; 76(1): 132-143, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34510371

RESUMO

Adenostemma lavenia (L.) Kuntze (Asteraceae) is widely distributed in tropical regions of East Asia, and both A. lavenia and A. madurense (DC) are distributed in Japan. In China and Taiwan, A. lavenia is used as a folk medicine for treating lung congestion, pneumonia, and hepatitis. However, neither phylogenic nor biochemical analysis of this plants has been performed to date. We have reported that the aqueous extract of Japanese A. lavenia contained high levels of ent-11α-hydroxy-15-oxo-kaur-16-en-19-oic acid (11αOH-KA; a kaurenoic acid), which is a potent anti-melanogenic compound. Comparison of chloroplast DNA sequences suggested that A. lavenia is originated from A. madurense. Analyses of kaurenoic acids revealed that Japanese A. lavenia and A. madurense contained high levels of 11αOH-KA and moderate levels of 11α,15OH-KA, while Taiwanese A. lavenia mainly contained 9,11αOH-KA. The diverse biological activities (downregulation of Tyr, tyrosinase, gene expression [anti-melanogenic] and iNOS, inducible nitric oxide synthase, gene expression [anti-inflammatory], and upregulation of HO-1, heme-oxygenase, gene expression [anti-oxidative]) were associated with 11αOH-KA and 9,11αOH-KA but not with 11α,15OH-KA. Additionally, 11αOH-KA and 9,11αOH-KA decreased Keap1 (Kelch-like ECH-associated protein 1) protein levels, which was accompanied by upregulation of protein level and transcriptional activity of Nrf2 (NF-E2-related factor-2) followed by HO-1 gene expression. 11αOH-KA and 9,11αOH-KA differ from 11α,15OH-KA in terms of the presence of a ketone (αß-unsaturated carbonyl group, a thiol modulator) at the 15th position; therefore, thiol moieties on the target proteins, including Keap1, may be important for the biological activities of 11αOH-KA and 9,11αOH-KA and A. lavenia extract.


Assuntos
Asteraceae , Fator 2 Relacionado a NF-E2 , Diterpenos , Heme Oxigenase-1/metabolismo , Japão , Proteína 1 Associada a ECH Semelhante a Kelch , Fator 2 Relacionado a NF-E2/metabolismo , Taiwan
10.
Biochem Biophys Res Commun ; 562: 50-54, 2021 07 12.
Artigo em Inglês | MEDLINE | ID: mdl-34034093

RESUMO

Mitochondria are eukaryotic organelles that consist of outer and inner bilayer membranes with a positive potential (H+) in the intermembrane space. This organelle plays an important role in ATP production and apoptosis. To observe the mitochondria in living cells, several fluorescent dyes (such as MitoTracker® [a standard mitochondrial imager] or rhodamine 123) have been developed. However, these reagents are unstable and exhibit a wide range of emission spectra, thereby hampering double staining results. Using recombinant DNA techniques, green or red fluorescent protein (GFP or RFP)-tagged proteins are now available for multi-color labeling of mitochondria. Here, we have discussed the development of the novel mitochondrial live imagers MitoMM1/2, derivatives of ATTO565; furthermore, MitoMM1/2 are sensitive to the membrane potential, resistant to detergents, and the fluorescence of MitoMM1/2 does not overlap with green fluorescence.


Assuntos
Corantes Fluorescentes/metabolismo , Mitocôndrias/metabolismo , Imagem Molecular , Corantes Fluorescentes/química , Proteínas de Fluorescência Verde/metabolismo , Células HEK293 , Humanos , Potencial da Membrana Mitocondrial , Mitofagia , Tetraspanina 30/metabolismo
11.
J Invest Dermatol ; 141(1): 105-113.e14, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-32470341

RESUMO

IL-33-activated group 2 innate lymphoid cells critically contribute to protease allergen-induced airway inflammation models. However, IL-33 is dispensable for a subcutaneous (s.c.) papain-induced skin inflammation model, suggesting distinct mechanisms between intranasal and s.c. sensitization. Here, we examined the role of IL-17A in the s.c. model. Papain-exposed skin produced IL-17A and an excess amount of a soluble decoy receptor for IL-33, with the latter being a possible reason for the independence of the s.c. model from IL-33. An IL-17A deficiency attenuated papain-induced skin eosinophilia and serum papain-specific IgE and IgG1 levels, whereas the s.c. administration of IL-17A with enzymatically inactive papain enhanced serum papain-specific IgE and IgG1 levels and T helper 2 development in draining lymph nodes in an IL-33-independent manner, suggesting IL-33-independent enhancement of papain-specific type 2 responses by IL-17A. The s.c. papain increased IL-17A+ γδ T cells in draining lymph nodes, approximately half of which were Vγ4+, as the majority of IL-17A+ cells, and increased Vγ5+ and Vγ4+ γδ T cells in the skin. Depletion of γδ TCR+ cells reduced T helper cytokine production in antigen-restimulated draining lymph node cells. These results suggest a novel role for IL-17A as an enhancer of skin eosinophilia and serum antigen-specific IgE production and for γδ T cells as an enhancer of T helper cell activation in the s.c. papain model.


Assuntos
Dermatite/imunologia , Eosinofilia/imunologia , Imunidade Inata , Interleucina-17/metabolismo , Interleucina-33/metabolismo , Papaína/administração & dosagem , Pele/patologia , Animais , Dermatite/metabolismo , Dermatite/patologia , Eosinofilia/metabolismo , Eosinofilia/patologia , Humanos , Injeções Subcutâneas , Pele/imunologia , Pele/metabolismo
12.
Heart Vessels ; 36(2): 200-210, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32902700

RESUMO

Although drug-eluting stents have improved clinical outcomes, percutaneous coronary intervention (PCI) for acute coronary syndrome (ACS) remains a challenging procedure in terms of thrombus management. A new-generation P2Y12 receptor inhibitor, prasugrel, provides more rapid and potent antiplatelet action compared with clopidogrel. Prasugrel achieved significant reduction of ischemic events compared with clopidogrel in ACS. The aim of this optical coherence tomography (OCT) study was to evaluate temporal changes in tissue prolapse after stenting under different antiplatelet regimens (aspirin plus prasugrel or clopidogrel) in ACS patients. A total of 119 ACS patients were randomized to either prasugrel or clopidogrel at the time of PCI. OCT analysis was available in 119 patients at baseline (just after stenting), 77 patients at 2 weeks, and 62 patients at 4 months after stenting. Cross-sectional analysis for every 1 mm was performed at in-stent and adjacent reference segment. Tissue prolapse area was calculated by lumen area minus stent area within the stented segment. Baseline patient and procedural characteristics were not different between the prasugrel and clopidogrel groups. Tissue prolapse area was significantly lower in the prasugrel compared with the clopidogrel group after stenting (0.24 ± 0.23 vs. 0.36 ± 0.23 mm2, p = 0.003) and at 2 weeks (0.11 ± 0.13 vs. 0.19 ± 0.16 mm2, p = 0.005). However, there was no significant difference at 4 months. In conclusion, our study suggests prasugrel was effective in reducing tissue prolapse in the super acute phase in ACS patients compared with clopidogrel. However, the effect of tissue prolapse reduction was not different up to 4 months follow-up.


Assuntos
Síndrome Coronariana Aguda/terapia , Vasos Coronários/diagnóstico por imagem , Stents Farmacológicos , Intervenção Coronária Percutânea , Inibidores da Agregação Plaquetária/uso terapêutico , Tomografia de Coerência Óptica/métodos , Síndrome Coronariana Aguda/diagnóstico , Idoso , Estudos Transversais , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Resultado do Tratamento
14.
Antioxidants (Basel) ; 9(8)2020 Aug 08.
Artigo em Inglês | MEDLINE | ID: mdl-32784463

RESUMO

BACKGROUND: The extract of Adenostemma lavenia (L.) O. Kuntze leaves has anti-inflammatory activities and is used as a folk medicine to treat patients with hepatitis and pneumonia in China and Taiwan. The diterpenoid ent-11α-hydroxy-15-oxo-kaur-16-en-19-oic acid (11αOH-KA) is the major ingredient in the extract and has wide-spectrum biological activities, such as antitumor and antimelanogenic activities, as well as anti-inflammatory activity. However, the physical and biological properties of this compound as an antioxidant or antiaging agent have not been reported yet. METHODS: In addition to in vitro assays, we monitored antioxidative and antiaging signals in Schizosaccharomyces pombe (yeast) and mouse melanoma B16F10 cells. RESULTS: A. lavenia water and chloroform fractions showed antioxidant properties in vitro. The A. lavenia extracts and 11αOH-KA conferred resistance to H2O2 to S. pombe and B16F10 cells and extended the yeast lifespan in a concentration-dependent manner. These materials maintained the yeast mitochondrial activity, even in a high-glucose medium, and induced an antioxidant gene program, the transcriptional factor pap1+ and its downstream ctt1+. Accordingly, 11αOH-KA activated the antioxidative transcription factor NF-E2-related factor 2, NRF2, the mammalian ortholog of pap1+, in B16F10 cells, which was accompanied by enhanced hemeoxygenase expression levels. These results suggest that 11αOH-KA and A. lavenia extracts may protect yeast and mammalian cells from oxidative stress and aging. Finally, we hope that these materials could be helpful in treating COVID-19 patients, because A. lavenia extracts and NRF2 activators have been reported to alleviate the symptoms of pneumonia in model animals.

15.
Biosci Biotechnol Biochem ; 84(4): 734-742, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31842701

RESUMO

scyllo-inositol dehydrogenase, isolated from Paracoccus laeviglucosivorans (Pl-sIDH), exhibits a broad substrate specificity: it oxidizes scyllo- and myo-inositols as well as L-glucose, converting L-glucose to L-glucono-1,5-lactone. Based on the crystal structures previously reported, Arg178 residue, located at the entry port of the catalytic site, seemed to be important for accepting substrates. Here, we report the role of Arg178 by using an alanine-substituted mutant for kinetic analysis as well as to determine the crystal structures. The wild-type Pl-sIDH exhibits the activity for scyllo-inositol most preferably followed by myo-inositol and L-glucose. On the contrary, the R178A mutant abolished the activities for both inositols, but remained active for L-glucose to the same extent as its wild-type. Based on the crystal structures of the mutant, the side chain of Asp191 flipped out of the substrate binding site. Therefore, Arg178 is important in positioning Asp191 correctly to exert its catalytic activities.Abbreviations: IDH: inositol dehydrogenase; LB: Luria-Bertani; kcat: catalyst rate constant; Km: Michaelis constant; NAD: nicotinamide dinucleotide; NADH: nicotinamide dinucleotide reduced form; PDB; Protein Data Bank; PDB entry: 6KTJ, 6KTK, 6KTL.


Assuntos
Substituição de Aminoácidos , Glucose/metabolismo , Inositol/metabolismo , Oxirredutases/metabolismo , Paracoccus/enzimologia , Cinética , Oxirredutases/isolamento & purificação , Conformação Proteica , Especificidade por Substrato
16.
J Vet Med Sci ; 82(2): 143-147, 2020 Feb 04.
Artigo em Inglês | MEDLINE | ID: mdl-31827015

RESUMO

In our previous studies, by simply inducing burn injuries on bullfrog hearts or partially exposing their surface to high-potassium (K+) solution, we could reproduce a ST segment elevation in the electrocardiogram (ECG), which is a characteristic finding in human ischemic heart disease. In the present study, using our burn-induced subepicardial injury model, we could additionally reproduce "reciprocal" ST segment changes for the first time in frog hearts, mimicking those observed in human acute myocardial infarction. Immunohistochemistry demonstrated markedly decreased Na+/K+-ATPase protein expression in the ventricular surface after the burn injury. The loss of this pump expression in injured cardiomyocytes was thought to be responsible for the creation of "currents of injury" and the subsequent ST segment changes observed in acute myocardial infarction.


Assuntos
Arritmias Cardíacas/patologia , Queimaduras , Modelos Animais de Doenças , Infarto do Miocárdio/fisiopatologia , Animais , Eletrocardiografia , Ventrículos do Coração/fisiopatologia , Masculino , Miócitos Cardíacos/metabolismo , Rana catesbeiana , ATPase Trocadora de Sódio-Potássio/metabolismo
17.
Ann Plast Surg ; 83(3): 305-307, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31232801

RESUMO

PURPOSE: The use of externalized jejunal monitoring flaps for jejunum transfers could be facilitative for the direct clinical assessment. Although this monitoring method would seem to be highly reliable, we modified this method and used mesentery only as a monitor to make it easy to manage the monitor more. METHODS: Between 2013 and 2018, 43 patients underwent vascularized jejunum transfer for reconstruction of laryngopharyngectomy using the externalized mesentery monitor. There were 39 men and 4 women, and patient ages ranged from 40 to 80 years (average, 66.6 years). The nursing staff monitored the externalized mesentery by using handheld Doppler ultrasonography every 2 hours for 7 days after surgery. RESULTS: Three patients had rather weak signal of handheld Doppler ultrasonography on the externalized mesentery monitors during operation, and handheld Doppler ultrasonography could not be applied. Of the remaining 40 patients using the externalized mesentery monitor with handheld Doppler ultrasonography, 39 had an uncomplicated postoperative period. In 1 patient, no signal of Doppler ultrasonography and lack of bleeding by pin prick from the monitor segment were noted in the immediate postoperative period, and revision of the vascular anastomosis was performed. Finally, the graft was salvaged. There was no case of infection in the monitoring flap or hypertrophic scar at the resected part of the flap. CONCLUSIONS: Using the externalized mesentery monitoring flaps, clinical monitoring by examining the exteriorized monitoring flap is possible, and only mesentery monitors were managed easily compared with jejunum monitoring flaps.


Assuntos
Retalhos de Tecido Biológico , Jejuno/transplante , Laringectomia , Faringectomia , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Mesentério/diagnóstico por imagem , Pessoa de Meia-Idade , Monitorização Fisiológica/métodos , Ultrassonografia Doppler
19.
Exp Ther Med ; 16(6): 5224-5226, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30546416

RESUMO

It is often difficult to correctly diagnose patients who present with dilation of the bile duct. Cholangiocarcinoma, primary sclerosing cholangitis (PSC) and immunoglobulin (Ig)G4-related sclerosing cholangitis must be considered as potential diagnoses for these cases. The current study presents a 73-year-old female patient who presented with a high fever and abdominal pain. Contrast-enhanced computed tomography and magnetic resonance cholangiopancreatography revealed stenosis and dilation of the intrahepatic bile duct without solid components. It was suspected that the patient had intrahepatic cholangiocarcinoma. A left liver lobectomy, cholecystectomy and distal gastrectomy combined with a D2 lymph node dissection were performed. A pathological examination of the liver revealed increased fibrosis in the stroma, irregular bile duct dilation and clusters of inflamed lymph cells. No carcinoma or IgG4-positive plasma cells were observed and the typical findings of PSC were not detected. Based on these clinical and pathological results, the diagnosis was idiopathic sclerosing cholangitis, which is particularly rare. It is often difficult to preoperatively differentiate between cholangiocarcinoma and benign bile duct stenosis.

20.
Proc Natl Acad Sci U S A ; 115(38): E8873-E8881, 2018 09 18.
Artigo em Inglês | MEDLINE | ID: mdl-30158171

RESUMO

The Ciona notochord has emerged as a simple and tractable in vivo model for tubulogenesis. Here, using a chemical genetics approach, we identified UTKO1 as a selective small molecule inhibitor of notochord tubulogenesis. We identified 14-3-3εa protein as a direct binding partner of UTKO1 and showed that 14-3-3εa knockdown leads to failure of notochord tubulogenesis. We found that UTKO1 prevents 14-3-3εa from interacting with ezrin/radixin/moesin (ERM), which is required for notochord tubulogenesis, suggesting that interactions between 14-3-3εa and ERM play a key role in regulating the early steps of tubulogenesis. Using live imaging, we found that, as lumens begin to open between neighboring cells, 14-3-3εa and ERM are highly colocalized at the basal cortex where they undergo cycles of accumulation and disappearance. Interestingly, the disappearance of 14-3-3εa and ERM during each cycle is tightly correlated with a transient flow of 14-3-3εa, ERM, myosin II, and other cytoplasmic elements from the basal surface toward the lumen-facing apical domain, which is often accompanied by visible changes in lumen architecture. Both pulsatile flow and lumen formation are abolished in larvae treated with UTKO1, in larvae depleted of either 14-3-3εa or ERM, or in larvae expressing a truncated form of 14-3-3εa that lacks the ability to interact with ERM. These results suggest that 14-3-3εa and ERM interact at the basal cortex to direct pulsatile basal accumulation and basal-apical transport of factors that are essential for lumen formation. We propose that similar mechanisms may underlie or may contribute to lumen formation in tubulogenesis in other systems.


Assuntos
Proteínas 14-3-3/fisiologia , Ciona intestinalis/embriologia , Células Endoteliais/fisiologia , Morfogênese/fisiologia , Proteínas 14-3-3/genética , Animais , Benzaldeídos/farmacologia , Ciona intestinalis/genética , Citoplasma/metabolismo , Proteínas do Citoesqueleto/genética , Proteínas do Citoesqueleto/metabolismo , Larva/crescimento & desenvolvimento , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Proteínas dos Microfilamentos/genética , Proteínas dos Microfilamentos/metabolismo , Morfogênese/efeitos dos fármacos , Morfogênese/genética , Morfolinos/genética , Miosina Tipo II/metabolismo , Notocorda/embriologia
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