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1.
Aging (Albany NY) ; 13(5): 6289-6297, 2021 03 12.
Artigo em Inglês | MEDLINE | ID: mdl-33711813

RESUMO

OBJECTIVES: To retrospectively evaluate the clinical and immunological characteristics of patients who died of COVID-19 and to identify patients at high risk of death at an early stage and reduce their mortality. RESULTS: Total white blood cell count, neutrophil count and C-reactive protein were significantly higher in patients who died of COVID-19 than those who recovered from it (p < 0.05), but the total lymphocyte count, CD4 + T cells, CD8 + T cells, B cells and natural killer cells were significantly lower when compared in the same groups. Multiple logistic regression analysis showed that increased D-dimer, decreased CD4 + T cells and increased neutrophils were risk factors for mortality. Further multiple COX regression demonstrated that neutrophil ≥ 5.27 × 109/L increased the risk of death in COVID-19 patients after adjustment for age and gender. However, CD4 + T cells ≥ 260/µL appeared to reduce the risk of death. CONCLUSION: SARS-CoV-2 infection led to a significant decrease of lymphocytes, and decreased CD4 + T cell count was a risk factor for COVID-19 patients to develop severe disease and death. METHODS: This study included 190 hospitalized COVID-19 patients from January 30, 2020 to March 4, 2020 in Wuhan, China, of whom 85 died and 105 recovered. Two researchers independently collected the clinical and laboratory data from electronic medical records.


Assuntos
/sangue , Linfócitos/imunologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Linfócitos B/imunologia , Proteína C-Reativa/análise , Proteína C-Reativa/imunologia , Linfócitos T CD4-Positivos/imunologia , /mortalidade , Feminino , Produtos de Degradação da Fibrina e do Fibrinogênio/análise , Produtos de Degradação da Fibrina e do Fibrinogênio/imunologia , Humanos , Células Matadoras Naturais/imunologia , Contagem de Linfócitos , Masculino , Pessoa de Meia-Idade , Neutrófilos/imunologia , Prognóstico , Estudos Retrospectivos , Fatores de Risco , /isolamento & purificação
2.
Mol Genet Genomics ; 295(5): 1227-1237, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32524299

RESUMO

The silk gland of the silkworm Bombyx mori is a specialized organ where silk proteins are efficiently synthesized under precise regulation that largely determines the properties of silk fibers. To understand the genes involved in the regulation of silk protein synthesis, considerable research has focused on the transcripts expressed in silk glands; however, the complete transcriptome profile of this organ has yet to be elucidated. Here, we report a full-length silk gland transcriptome obtained by PacBio single-molecule long-read sequencing technology. In total, 11,697 non-redundant transcripts were identified in mixed samples of silk glands dissected from larvae at five developmental stages. When compared with the published reference, the full-length transcripts optimized the structures of 3002 known genes, and a total of 9061 novel transcripts with an average length of 2171 bp were detected. Among these, 1403 (15.5%) novel transcripts were computationally revealed to be lncRNAs, 8135 (89.8%) novel transcripts were annotated to different protein and nucleotide databases, and 5655 (62.4%) novel transcripts were predicted to have complete ORFs. Furthermore, we found 1867 alternative splicing events, 2529 alternative polyadenylation events, 784 fusion events and 6596 SSRs. This study provides a comprehensive set of reference transcripts and greatly revises and expands the available silkworm transcript data. In addition, these data will be very useful for studying the regulatory mechanisms of silk protein synthesis.


Assuntos
Bombyx/crescimento & desenvolvimento , Perfilação da Expressão Gênica/métodos , Seda/genética , Imagem Individual de Molécula/métodos , Processamento Alternativo , Animais , Bombyx/genética , Regulação da Expressão Gênica no Desenvolvimento , Sequenciamento de Nucleotídeos em Larga Escala , Proteínas de Insetos/genética , Fases de Leitura Aberta , Poliadenilação , RNA Longo não Codificante/genética , Sequenciamento Completo do Exoma
3.
Zhong Nan Da Xue Xue Bao Yi Xue Ban ; 45(3): 334-337, 2020 Mar 28.
Artigo em Inglês, Chinês | MEDLINE | ID: mdl-32386027

RESUMO

The current outbreak of coronavirus disease 2019 (COVID-19) in Wuhan, China, has posed significant threats to international health. By Feb. 20, 2020, 74 576 cases have been confirmed and over 2 118 deaths have reported in the Chinese mainland. Chinese administrations have carried out immediate and prompt measures to stop the spread of the virus. Wuhan city has been shut down since Jan. 23, and more than 30 thousand medical workers have been recruited to Hubei province. Two temporary hospitals were constructed to treat severe pneumonia patients, and 15 mobile cabin hospitals were built to treat mild pneumonia cases. Significant improvement regarding the pathogenesis, epidemiology, and diagnosis and therapy for the COVID-19 has been achieved to stop the spread of the epidemics.


Assuntos
Betacoronavirus , Infecções por Coronavirus/epidemiologia , Pneumonia Viral/epidemiologia , China , Epidemias , Humanos , Pandemias
4.
Toxicol In Vitro ; 66: 104881, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32360864

RESUMO

It is critical to understand the risk factors responsible for the tumorigenesis and progression of nasopharyngeal carcinoma (NPC). Bisphenol A (BPA) can regulate the estrogenic signals to modulate cancer progression, while its roles in NC were not investigated. Our present study revealed that the BPA can increase proliferation and migration of NPC cells while decrease the chemosensitivity to doxorubicin (Dox). The inhibitor of GSK-3ß/ß-catenin (LiCl) can restore BPA-induced cell proliferation of NPC cells, which is due to that BPA can decrease phosphorylation while increase expression and nucleus localization of ß-catenin. Mechanistically, BPA can increase the mRNA stability of ß-catenin (encoded by CTNNB1) via suppressing the expression of miR-214-3p, which can direct target the 3'UTR of ß-catenin mRNA. Further, BPA can decrease phosphorylation of ß-catenin via repressing the expression of CK1α. Collectively, our data showed that BPA can trigger the proliferation and malignancy of NPC cells via activation of Wnt/ß-catenin pathway. It indicated that body accumulation and inhalation exposure of BPA might be a risk factor for NPC development.

5.
Chem Asian J ; 15(7): 1147-1155, 2020 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-32030909

RESUMO

When the 1,1,4,4-tetraanilinobutadiene skeleton is attached with two halogenated aryl units at the 2,3-positions, they undergo facile two-electron oxidation to give stable dicationic dyes which exhibit a near-infrared (NIR) absorption whereas the neutral dienes show only pale color. Therefore, a distinct electrochromic response with an absorption change in the NIR region is achieved, which is attracting considerable recent attention from the viewpoint of bioimaging. Herein, we demonstrate that the redox potentials of the 1,1,4,4-tetraanilinobutadiene can be precisely controlled by the donating properties of the amino group on the aniline unit as well as the number of halogen atoms on the aryl units at 2,3-positions on the butadiene. In contrast, the NIR absorption bands mainly depend on the number of halogen atoms irrespective to the donating properties of aniline unit. Thus, the hexaarylbutadiene skeleton is proven to be a versatile scaffold to develop less-explored organic NIR electrochromic materials, whose redox and spectroscopic properties can be finely tuned by modifying/attaching the proper substituents.

6.
Transgenic Res ; 29(2): 243-251, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-31960249

RESUMO

The cytoplasmic actin gene Actin4 (A4) in silkworm (Bombyx mori) was isolated 20 years ago and has a distal promoter upstream of the first exon and a proximal promoter within the first intron; however, how the promoter regulates gene expression has yet to be fully elucidated. Here, we characterized the function and expression of the proximal promoter (named A4IP) by analyzing transgenic Gal4/UAS silkworms, A4IP-Gal4/UAS-EGFP. We demonstrated that A4IP drives the expression of Gal4 and thereby activates UAS-linked EGFP in transgenic silkworms beginning in day-3 embryos through adults. Further detection revealed that EGFP was expressed at a low level in tissues including the trachea, fat body and midgut but was highly expressed in the wing disks/wings and inner epidermis of transgenic silkworms. No EGFP signals were detected in other tissues by western blot assay. Interestingly, EGFP fluorescence had a spot-like distribution on the epidermis of transgenic larvae. These observations are quite different from those in transgenic silkworms driven by the promoter of Actin3 (A3), another cytoplasmic actin gene in B. mori. These findings reveal the expression profiles of the A4IP promoter and provide new insights into the regulatory mechanism of cytoplasmic actin genes in silkworms.

7.
Nat Commun ; 11(1): 446, 2020 01 23.
Artigo em Inglês | MEDLINE | ID: mdl-31974383

RESUMO

Afterglow luminescent probes with high signal-to-background ratio show promise for in vivo imaging; however, such probes that can be selectively delivered into target sites and switch on afterglow luminescence remain limited. We optimize an organic electrochromic material and integrate it into near-infrared (NIR) photosensitizer (silicon 2,3-naphthalocyanine bis(trihexylsilyloxide) and (poly[2-methoxy-5-(2-ethylhexyloxy)-1,4-phenylenevinylene]) containing nanoparticles, developing an H2S-activatable NIR afterglow probe (F12+-ANP). F12+-ANP displays a fast reaction rate (1563 ± 141 M-1 s-1) and large afterglow turn-on ratio (~122-fold) toward H2S, enabling high-sensitivity and -specificity measurement of H2S concentration in bloods from healthy persons, hepatic or colorectal cancer patients. We further construct a hepatic-tumor-targeting and H2S-activatable afterglow probe (F12+-ANP-Gal) for noninvasive, real-time imaging of tiny subcutaneous HepG2 tumors (<3 mm in diameter) and orthotopic liver tumors in mice. Strikingly, F12+-ANP-Gal accurately delineates tumor margins in excised hepatic cancer specimens, which may facilitate intraoperative guidance of hepatic cancer surgery.


Assuntos
Carcinoma Hepatocelular/diagnóstico por imagem , Sulfeto de Hidrogênio/análise , Neoplasias Hepáticas/diagnóstico por imagem , Substâncias Luminescentes/química , Imagem Molecular/métodos , Animais , Neoplasias Colorretais/sangue , Cistationina beta-Sintase/análise , Cistationina beta-Sintase/metabolismo , Cistationina gama-Liase/análise , Cistationina gama-Liase/metabolismo , Células Hep G2 , Humanos , Sulfeto de Hidrogênio/sangue , Sulfeto de Hidrogênio/química , Neoplasias Hepáticas/sangue , Neoplasias Hepáticas Experimentais/diagnóstico por imagem , Substâncias Luminescentes/síntese química , Camundongos Endogâmicos BALB C , Nanopartículas/química , Fármacos Fotossensibilizantes/química , Polímeros/química , Compostos de Vinila/química , Ensaios Antitumorais Modelo de Xenoenxerto
8.
Biochem Biophys Res Commun ; 522(2): 492-498, 2020 02 05.
Artigo em Inglês | MEDLINE | ID: mdl-31780262

RESUMO

Sericin, produced in the middle silk gland (MSG) of silkworms, is a group of glue proteins that coat and cement silk fibers. Several genes are known to encode sericin, but their spatiotemporal regulation has yet to be fully elucidated. Here, we report in detail the expression profiles of the promoters of two major sericin-coding genes, Sericin 1 (Ser1)and Sericin 3 (Ser3), by analyzing Gal4/UAS transgenic silkworms. We found that UAS-linked EGFP fluorescence in transgenic silkworms driven by Ser1-Gal4was detected in only the R3, R4 and R5 regions of MSG starting inday-3 fifth-instar larvae and was continuously expressed until silk gland degradation. In transgenic silkworms driven by Ser3-Gal4, EGFP fluorescence was detected at a low level in the R2 region of MSG since the last day of fifth-instar larvae, and the expression increased during the wandering stages and was continuously detected until silk gland degradation. The molecular detection of EGFP expression in each of the Gal4/UAS transgenic silkworms was consistent with fluorescence observations. These findings reveal clear differences in the regulatory characteristics of the promoters of Ser1and Ser3 and provide new insights into the regulatory mechanism of the expression of sericin-coding genes.


Assuntos
Bombyx/genética , Regiões Promotoras Genéticas , Sericinas/genética , Animais , Animais Geneticamente Modificados , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Fluorescência Verde/metabolismo , Larva/genética , Pupa/genética , Sericinas/metabolismo
9.
Environ Entomol ; 49(1): 21-32, 2020 02 17.
Artigo em Inglês | MEDLINE | ID: mdl-31782953

RESUMO

The lignocellulosic digestive symbiosis in termites is a dynamic survival adaptation system. While the contribution of hereditary and habitat factors to the development of the symbiotic bacterial community of termites had been confirmed, the manner in which these factors affect functional synergism among different bacterial lineages has still not been fully elucidated. Therefore, the 16S rRNA gene libraries of Odontotermes formosanus Shiraki (Blattodea: Termitidae) and Coptotermes formosanus Shiraki (Blattodea: Rhinotermitidae) sampled from sugarcane fields (high sugar) or pine tree forests (no free sugar) were sequenced. The results verify that the prokaryotic community structures of termites could be significantly reshaped by native dietary isolation within a species. Although the most dominant phyla are convergent in all samples, their relative abundances in these two termite species exhibited a reverse variation pattern when the termite hosts were fed on the high-sugar diet. Furthermore, we showed that the taxonomic composition of the dominant phyla at the family or genus level differentiate depending on the diet and the host phylogeny. We hypothesize that the flexible bacterial assemblages at low taxonomic level might exert variable functional collaboration to accommodate to high-sugar diet. In addition, the functional predictions of Tax4Fun suggest a stable metabolic functional structure of the microbial communities of the termites in both different diet habitats and taxonomy. We propose that the symbiotic bacterial community in different host termites developed a different functional synergistic pattern, which may be essential to maintain the stability of the overall metabolic function for the survival of termites.


Assuntos
Baratas , Isópteros/genética , Animais , Dieta , RNA Ribossômico 16S , Análise de Sequência de DNA , Açúcares
10.
Dev Neurorehabil ; 23(3): 176-184, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31088323

RESUMO

PURPOSE: To evaluate the influence of accuracy constraints on functional symmetric bimanual coordination for children with Unilateral Spastic Cerebral Palsy (USCP). METHODS: Ten children with USCP (average age: 9.6; MACS levels: I-II), ten typically developing children, and ten adults lifted a tray with a water bottle on top. Two accuracy constraints of handle size and cap condition were manipulated. RESULTS: Children with USCP exhibited greater bilateral asymmetry in hand vertical position, timing, upper arm, and elbow control than other groups. Smaller handle decreased bilateral timing differences at lift onset and offset, and decreased bilateral elbow asymmetry at reach and lift offset. Without a cap (accuracy constraint), they showed greater trunk involvement, and less bilateral vertical position and lift offset timing differences (all p < .05). CONCLUSIONS: Children with USCP showed impaired symmetric bimanual coordination. Higher accuracy constraints improved some bimanual spatial and temporal control. Therefore, task accuracy constraints should be manipulated carefully for training.


Assuntos
Paralisia Cerebral/fisiopatologia , Mãos/fisiopatologia , Destreza Motora , Criança , Feminino , Humanos , Masculino , Movimento
11.
Prev Med Rep ; 16: 101010, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31799106

RESUMO

Annually, about 16,500 HPV-associated cancers occur in the US among men. Data regarding HPV vaccine uptake among men based on nativity status (i.e., US-versus foreign-born) is limited, yet potentially important for informing interventions. We assessed differences in HPV vaccine uptake by nativity status among men aged 18-34 years in the US. The 2014-2017 National Health Interview Survey was examined for men, aged 18-34 years (n = 14,056). HPV vaccine initiation was defined as receipt of at least one dose of the vaccine and completion as receipt of three doses. Weighted, multivariable binary logistic regression models were used to assess the association between nativity status and HPV vaccine uptake, adjusting for demographic, socioeconomic, and healthcare factors. Analyses were performed in July 2018. Overall, 17% of men self-identified as foreign-born, 9.9% of men had initiated the HPV vaccine, and 3.3% had completed the HPV vaccine. Among foreign-born men, Asians had the highest HPV vaccination rates whereas those from Indian subcontinental region had the lowest rates. After accounting for demographic, socioeconomic, and healthcare factors, compared to US-born men, foreign-born men were 46% (adjusted odds ratio = 0.54; 95% CI = 0.39-0.72) less likely to initiate the HPV vaccine but there was no difference between the two groups in terms of vaccine completion. We found that HPV vaccine uptake among men was very low overall, and foreign-born men had lower initiation compared to US-born men. Public health interventions which improve HPV vaccination need to be developed for all men, irrespective of nativity status.

12.
Sensors (Basel) ; 20(1)2019 Dec 23.
Artigo em Inglês | MEDLINE | ID: mdl-31878080

RESUMO

The mechanical dithered ring laser gyro (RLG) effectively overcomes the lock-in effect and ensures the sensitive accuracy of the low angular rate for the gyro. However, in the inertial measurement unit (IMU) system, the dither excitation of three RLGs causes the coupled vibration of the IMU structure, which could seriously limit the measuring accuracy of RLGs. In this paper, the vibration frequency response characteristic of laser gyro IMU is taken as the focus point, and the method of multi-rigid body dynamics is used to establish the dynamic model of IMU suitable for vibration frequency response analysis. On the basis of the model, the multi-degree-of-freedom coupling vibration of IMU with the gyro dither excitation is clearly described. A new IMU dynamic decoupling optimization method is proposed to minimize the coupled vibration frequency response, and compared with the previous optimal design method. The prototype experimental test results show that the coupled vibration of IMU is restrained more effectively by the proposed new method than by the previous optimal design method. Finally, on the basis of this new method, the measuring accuracy of the RLGs in the IMU system is improved, which is quite useful for practical engineering application.

13.
Transgenic Res ; 28(5-6): 627-636, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31564030

RESUMO

The silkworm Bombyx mori is a valuable insect that synthesizes bulk amounts of fibroin protein in its posterior silk gland (PSG) and weaves these proteins into silk cocoons. The mechanism by which the fibroin protein is efficiently synthesized and precisely regulated is an important aspect that has yet to be fully elucidated. Here, we describe the regulatory characteristics of the promoters of fibroin protein-encoding genes, namely, fibroin heavy chain (fibH) and fibroin light chain (fibL), using an optimized Gal4/UAS binary system. We found that (1) UAS-linked enhanced green fluorescent protein (EGFP) was effectively activated in the PSGs of Gal4/UAS transgenic silkworms, and fluorescence was continuously detected in the PSGs after complete formation of silk glands. (2) In the PSGs of fourth- and fifth-instar larvae of transgenic silkworms driven by fibL-Gal4 (LG4) or fibH-Gal4 (HG4), EGFP mRNA was detected in only day-3 to day-6 fifth-instar larvae, while the EGFP protein could be detected at each day of both larval stages. (3) High-level expression of Gal4 and UAS-linked EGFP caused a delay in PSG degradation in Gal4/UAS transgenic silkworms. (4) At the early pupal stage, EGFP fluorescence was also detected in fat bodies of Gal4/UAS transgenic silkworms, indicating that the PSG-specific EGFP was transported into fat bodies during PSG degeneration; however, the underlying mechanism needs to be further elucidated. This study provides a modified Gal4/UAS system used for efficient tissue-specific expression of target genes in the PSGs of silkworms and provides new insights into the regulatory characteristics of the promoters of key fibroin protein-encoding genes.


Assuntos
Animais Geneticamente Modificados/genética , Bombyx/genética , Fibroínas/genética , Proteínas de Insetos/genética , Animais , Fibroínas/biossíntese , Proteínas de Fluorescência Verde/genética , Larva/genética , Regiões Promotoras Genéticas/genética , Pupa/genética , Seda/genética , Fatores de Transcrição
14.
Curr Med Sci ; 39(3): 415-418, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31209812

RESUMO

The colon is an alternative graft organ for esophageal reconstruction. The present study reviewed our experience with the colon interposition for esophageal replacement following corrosive ingestion, to evaluate the outcomes of colon interposition based on our surgical experience. The clinical data of 119 patients who underwent colon interposition for esophageal replacement from January 2005 to March 2017 were retrospectively analyzed. The routes of the colon interposition were retrosternal in 119 (100%). The median operative time was 390 min (range: 290-610 min) and the median blood loss was 615 mL (range: 270-2500 mL). Of these 119 patients, the cervical anastomosis was performed at the hypopharynx (n=20, 16.8%), the larynx (n=3, 2.5%), and the cervical esophagus (n=96, 80.7%). Five patients experienced cervical anastomotic leakage (4 cases for esophagus-colon, and one for hypopharynx-colon). One patient experienced wound infection of the abdominal wall. Three patients had injury of recurrent laryngeal nerve and hoarseness. Three patients had stress ulcer with bleeding and treated with octreotide. Two patients suffered from incomplete intestinal obstruction. The postoperative follow-up was made for 12 months in all patients and all of them were alive. In conclusion, The colon is well-suited for esophageal reconstruction. The selection of the colon graft should be flexible and be based on the inspection of blood supply and the length needed. We must therefore make every effort to reduce the number of postoperative complications, and improve the quality of life for patients.


Assuntos
Colo/cirurgia , Estenose Esofágica/cirurgia , Esôfago/cirurgia , Procedimentos Cirúrgicos Reconstrutivos/métodos , Transplante Autólogo/métodos , Adolescente , Adulto , Idoso , Anastomose Cirúrgica/métodos , Criança , Pré-Escolar , Colo/fisiologia , Traumatismos dos Nervos Cranianos/diagnóstico , Traumatismos dos Nervos Cranianos/etiologia , Traumatismos dos Nervos Cranianos/fisiopatologia , Estenose Esofágica/fisiopatologia , Esôfago/fisiopatologia , Feminino , Seguimentos , Hemorragia/diagnóstico , Hemorragia/etiologia , Hemorragia/fisiopatologia , Humanos , Obstrução Intestinal/diagnóstico , Obstrução Intestinal/etiologia , Obstrução Intestinal/fisiopatologia , Nervos Laríngeos/cirurgia , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/diagnóstico , Complicações Pós-Operatórias/fisiopatologia , Resultado do Tratamento
15.
J Pharmacol Sci ; 140(1): 14-19, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31113729

RESUMO

Secreted frizzled-related protein (Sfrp) 4 is a protein that involve in cardiac development and several cardiovascular diseases. However, the effect of Sfrp4 in mediating myocardial ischemia/reperfusion injury remains unknown. In this work, adenoviral (Ad)-shSfrp4 adenoviruses was used to knockdown of Sfrp4 in myocardium to examine the role of Sfrp4 in mediating myocardial I/R injury. Knockdown of Sfrp4 in mice attenuated myocardial I/R injury, as indicated by the decrease levels of lactate dehydrogenase and creatine kinase, and increment of ventricular function following I/R injury. Besides, knockdown of Sfrp4 led to a reduction in Bax, active caspase 3, and increase Bcl-2 and c-Myc in cardiac tissue. Knockdown of Sfrp4 lost its protection against I/R injury in mice infected with Ad-dn-AKT. In conclusion, knockdown of Sfrp4 in myocardium attenuated myocardial ischemia and reperfusion injury by AKT signaling pathway.


Assuntos
Apoptose/genética , Técnicas de Silenciamento de Genes , Traumatismo por Reperfusão Miocárdica/genética , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Proteínas Proto-Oncogênicas/genética , Proteínas Proto-Oncogênicas/fisiologia , Animais , Masculino , Camundongos Endogâmicos C57BL , Miocárdio/metabolismo , Proteínas Proto-Oncogênicas c-akt , Transdução de Sinais/genética
16.
Gene ; 646: 39-46, 2018 Mar 10.
Artigo em Inglês | MEDLINE | ID: mdl-29289608

RESUMO

Yorkie (Yki), the Drosophila homolog of vertebrate yes-associated protein (YAP), is a key effector of the Hippo pathway, which modulates organ size via the transcriptional regulation of downstream targets involved in cell proliferation and survival. YAP has been shown to be expressed as multiple splicing isoforms in mammals, but thus far, no evidence of alternatively spliced Yki isoforms has been reported in insects. Here, we confirmed that the Yki protein of the silkworm Bombyx mori, BmYki, is transcribed in the silk gland into at least four splicing isoforms, named BmY1329, BmY1314, BmY1188, and BmY1173. Further analysis revealed that BmY1329 and BmY1314 each contain two WW domains, whereas BmY1188 and BmY1173 each contain only one WW domain. Each BmYki isoform functions in regulating expression of Yki target genes in cultured B. mori embryonic cells, and exhibits a few different effects on the expression of Yki targets. Interestingly, the expression of silk fibroin protein genes could also be influenced by each of the BmYki isoforms, suggesting that BmYki is involved in the regulation of silk protein-coding genes. This study provides novel insights into the role of BmYki. The contribution of each BmYki isoform to the modulation of gene expression will be of great interest for further study.


Assuntos
Processamento Alternativo , Bombyx/genética , Transativadores/genética , Transativadores/metabolismo , Animais , Bombyx/crescimento & desenvolvimento , Bombyx/metabolismo , Células Cultivadas , Células-Tronco Embrionárias , Regulação da Expressão Gênica , Proteínas de Insetos/química , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Domínios Proteicos , Análise de Sequência de DNA , Seda/genética , Seda/metabolismo , Distribuição Tecidual , Transativadores/química , Transcrição Genética
17.
Transgenic Res ; 26(6): 807-815, 2017 12.
Artigo em Inglês | MEDLINE | ID: mdl-28952064

RESUMO

Diapause is a state of developmental arrest that is most often observed in arthropods, especially insects. The domesticated silkworm, Bombyx mori, is a typical insect that enters diapause at an early embryonic stage. Previous studies have revealed that the diapause hormone (DH) signaling molecules, especially the core members DH and DH receptor 1 (DHR1), are crucial for the determination of embryonic diapause in diapause silkworm strains. However, whether they function in non-diapause silkworm strains remains largely unknown. Here, we generated two transgenic lines overexpressing DH or DHR1 genes in a non-diapause silkworm strain, Nistari. Our results showed that developmental expression patterns of DH and DHR1 are quite similar in transgenic silkworms: both genes are highly expressed in the mid to late stages of pupae and are most highly expressed in day-6 pupae but are expressed at very low levels in other developmental stages. Moreover, the overexpression of DH or DHR1 can affect the expression of diapause-related genes but is not sufficient to induce embryonic diapause in their offspring. This study provides new insights into the function of DH and DHR1 in a non-diapause silkworm strain.


Assuntos
Bombyx/genética , Proteínas de Insetos/genética , Neuropeptídeos/genética , Animais , Animais Geneticamente Modificados , Bombyx/fisiologia , Feminino , Regulação da Expressão Gênica , Fenótipo
18.
PLoS One ; 12(8): e0182690, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28793345

RESUMO

The transcriptional coactivator Yorkie(Yki), is a critical downstream effector of the Hippo(Hpo) signaling pathway that controls organ size through the regulation of cell proliferation and apoptosis. During the past ten years the biological function of Yki has been studied extensively in Drosophila and a few other insects, however, little is known about it in the silkworm, Bombyx mori, a major research model of lepidopteran insect. Here, we describe the isolation, characterization and expression of the B. mori Yki ortholog, BmYki. The coding sequence of the BmYki was 1314 bp in length, encoding a protein of 437 amino acids containing two conserved WW domains. BmYki transcripts were ubiquitous but not abundant in all detected tissues and developmental stages. Comparatively, it was expressed at pretty high level in silk glands and at the stage of fifth-instar day-3 larvae. Overexpression of BmYki in cultured B. mori embryonic cells significantly promoted transcription of genes associated with cell proliferation and apoptosis, indicating that BmYki functions in the regulation of organ growth-related biological processes. Interestingly, transcription of silk protein-coding genes and transcription factors regulating the synthesis of silk proteins was downregulated remarkably, suggesting that BmYki was involved in the regulation of silk protein synthesis. This study provides new insights into the role of BmYki in Hpo pathway regulation in silkworm.


Assuntos
Bombyx/genética , Genes de Insetos/genética , Animais , Apoptose/genética , Bombyx/crescimento & desenvolvimento , Proliferação de Células/genética , Clonagem Molecular , Perfilação da Expressão Gênica , Larva/crescimento & desenvolvimento , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Transativadores/genética
19.
ACS Omega ; 2(5): 2034-2040, 2017 May 31.
Artigo em Inglês | MEDLINE | ID: mdl-31457557

RESUMO

We present an investigation of the photochemistry of aluminum nanoparticles (Al NPs) capped with poly(methyl methacrylate) (PMMA). Powder X-ray diffraction and Fourier transform infrared spectroscopy with total attenuated reflection confirm the presence of crystalline aluminum cores and the PMMA cap and allow us to confirm the latter's photodegradation upon exposure to UV light. The PMMA-Al NPs were also characterized by differential scanning calorimetry coupled with thermogravimetric analysis to study the thermal profiles for polymer combustion and metal oxidation exotherms. Transmission electron microscopy confirms that the Al NPs, around 36 nm in diameter, are embedded in the PMMA matrix. Following UV irradiation, the PMMA-Al NPs react considerably faster with alkaline solutions, compared with unphotolyzed samples. Photoactivation of the nanocomposite induces partial decomposition of the PMMA capping layer, exposing the underlying reactive metal cores to the surrounding environment and accelerating its redox reactivity. Photolysis times of 1, 6, 24, and 52 h were investigated to establish a minimum UV exposure time for the activation of the PMMA-Al NPs toward hydrolytic hydrogen gas generation.

20.
Anal Chim Acta ; 950: 178-183, 2017 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-27916123

RESUMO

A mitochondria-targeted fluorescence probe (CPBT) for ratiometric detection of endogenous hypochlorite in the living cells was developed. CPBT could detect hypochlorite with high selectivity and sensitivity in a ratiometric manner based on FRET mechanism. In absence of hypochlorite, when CPBT was excited with absorption maximum wavelength of the donor moiety, it showed the emission of acceptor moiety because of FRET process. However, in the presence of hypochlorite, the reaction of CC double bond with hypochlorite interrupted the conjugation system resulting in the inhibition of FRET process and the emission of the donor moiety. The two well-resolved emission bands can ensure accurate detection of hypochlorite. A good linear relationship between the fluorescence intensity ratios of the two emissions and the ClO- concentrations in the range from 41.8 nM (detection limit) to 12.5 µM was established. Importantly, CPBT could localize mainly in the mitochondria of RAW264.7 cells. CPBT was successfully used to fluorescence ratiometric imaging of endogenous hypochlorite in RAW264.7 cells.


Assuntos
Corantes Fluorescentes , Ácido Hipocloroso/análise , Mitocôndrias/química , Animais , Transferência Ressonante de Energia de Fluorescência , Camundongos , Microscopia de Fluorescência , Células RAW 264.7
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