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1.
Int J Mol Sci ; 22(19)2021 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-34639064

RESUMO

The signal transduction of the equine lutropin/choriogonadotropin receptor (eLH/CGR) is unclear in naturally occurring activating/inactivating mutants of this receptor, which plays an important role in reproductive physiology. We undertook the present study to determine whether conserved structurally related mutations in eLH/CGR exhibit similar mechanisms of signal transduction. We constructed four constitutively activating mutants (M398T, L457R, D564G, and D578Y) and three inactivating mutants (D405N, R464H, and Y546F); measured cyclic adenosine monophosphate (cAMP) accumulation via homogeneous time-resolved fluorescence assays in Chinese hamster ovary cells; and investigated cell-surface receptor loss using an enzyme-linked immunosorbent assay in human embryonic kidney 293 cells. The eLH/CGR-L457R-, -D564G-, and -D578Y-expressing cells exhibited 16.9-, 16.4-, and 11.2-fold increases in basal cAMP response, respectively. The eLH/CGR-D405N- and R464H-expressing cells presented a completely impaired signal transduction, whereas the Y546F-expressing cells exhibited a small increase in cAMP response. The cell-surface receptor loss was 1.4- to 2.4-fold greater in the activating-mutant-expressing cells than in wild-type eLH/CGR-expressing cells, but was completely impaired in the D405N- and Y546F-expressing cells, despite treatment with a high concentration of agonist. In summary, the state of activation of eLH/CGR influenced agonist-induced cell-surface receptor loss, which was directly related to the signal transduction of constitutively activating mutants.

2.
BMC Biotechnol ; 21(1): 52, 2021 09 05.
Artigo em Inglês | MEDLINE | ID: mdl-34482828

RESUMO

BACKGROUND: Equine chorionic gonadotropin (eCG), which comprises highly glycosylated α-subunit and ß-subunit, is a unique member of the glycoprotein hormone family as it elicits both follicle-stimulating hormone (FSH)-like and luteinizing hormone (LH)-like responses in non-equid species. To examine the biological function of glycosylated sites in eCG, the following glycosylation site mutants were constructed: eCGß/αΔ56, substitution of Asn56 of α-subunit with Gln; eCGß-D/α, deletion of the O-linked glycosylation site at the carboxyl-terminal peptide (CTP) region of the ß-subunit; eCGß-D/αΔ56, double mutant. The recombinant eCG (rec-eCG) mutants were expressed in Chinese hamster ovary suspension (CHO-S) cells. The FSH-like and LH-like activities of the mutants were examined using CHO-K1 cells expressing rat lutropin/CG receptor (rLH/CGR) and rat FSH receptor (rFSHR). RESULTS: Both rec-eCGß/α and rec-eCGß/αΔ56 were efficiently secreted into the CHO-S cell culture medium on day 1 post-transfection. However, the secretion of eCGß-D/α and eCGß-D/αΔ56, which lack approximately 12 O-linked glycosylation sites, was slightly delayed. The expression levels of all mutants were similar (200-250 mIU/mL) from days 3 to 7 post-transfection. The molecular weight of rec-eCGß/α, rec-eCGß/αΔ56 and rec-eCG ß-D/α were in the ranges of 40-45, 37-42, and 34-36 kDa, respectively. Treatment with peptide-N-glycanase F markedly decreased the molecular weight to approximately 5-10 kDa. Rec-eCGß/αΔ56 exhibited markedly downregulated LH-like activity. The signal transduction activity of both double mutants was completely impaired. This indicated that the glycosylation site at Asn56 of the α-subunit plays a pivotal role in the LH-like activity of eCG. Similarly, the FSH-like activity of the mutants was markedly downregulated. eCGß-D/α exhibited markedly downregulated LH-like and FSH-like activities. CONCLUSIONS: Rec-eCGß/α exhibits potent biological activity in cells expressing rLH/CGR and rFSHR. The findings of this study suggest that the LH-like and FSH-like activities of eCG are regulated by the N-linked glycosylation site at Asn56 of the eCG α-subunit and/or by the O-linked glycosylation sites of the eCG ß-subunit. These findings improved our understanding of the mechanisms underlying both LH-like and FSH-like activities of eCG.

3.
Anim Biosci ; 2021 Aug 24.
Artigo em Inglês | MEDLINE | ID: mdl-34474536

RESUMO

Objective: Follicle-stimulating hormone (FSH) is the central hormone involved in mammalian reproduction, maturation at puberty, and gamete production that mediates its function by control of follicle growth and function. The present study investigated the mutations involved in the regulation of FSH receptor (FSHR) activation. Methods: We analyzed seven naturally-occurring mutations that were previously reported in human FSHR (hFSHR), in the context of equine FSHR (eFSHR); these include one constitutively activation variant, one allelic variant, and five inactivating variants. These mutations were introduced into wild-type eFSHR (eFSHR-wt) sequence to generate mutants that were designated as eFSHR-D566G, -A306T, -A189V, -N191I, -R572C, -A574V, and -R633H. Mutants were transfected into PathHunter EA-parental CHO-K1 cells expressing ß-arrestin. The biological function of mutants was analyzed by quantitating cAMP accumulation in cells incubated with increasing concentrations of FSH. Results: Cells expressing eFSHR-D566G exhibited an 8.6-fold increase in basal cAMP response, as compared to that in eFSHR-wt. The allelic variation mutant eFSHR-A306T was not found to affect the basal cAMP response or EC50 levels. On the other hand, eFSHR-D566G and eFSHR-A306T displayed a 1.5- and 1.4-fold increase in the maximal response, respectively. Signal transduction was found to be completely impaired in case of the inactivating mutants eFSHR-A189V, -R572C, and -A574V. When compared with eFSHR-wt, eFSHR-N191I displayed a 5.4-fold decrease in the EC50 levels (3910 ng/mL) and a 2.3-fold decrease in the maximal response. In contrast, cells expressing eFSHR-R633H displayed in a similar manner to that of the cells expressing the eFSHR-wt on signal transduction and maximal response. Conclusion: The activating mutant eFSHR-D566G greatly enhanced the signal transduction in response to FSH, in the absence of agonist treatment. We suggest that the state of activation of the eFSHR can modulate its basal cAMP accumulation.

4.
Int J Mol Sci ; 22(15)2021 Jul 22.
Artigo em Inglês | MEDLINE | ID: mdl-34360584

RESUMO

Trichostatin A (TSA) is a representative histone deacetylase (HDAC) inhibitor that modulates epigenetic gene expression by regulation of chromatin remodeling in cells. To investigate whether the regulation of chromatin de-condensation by TSA can affect the increase in the efficiency of Cas9 protein-gRNA ribonucleoprotein (RNP) indel formation from plant cells, genome editing efficiency using lettuce and tobacco protoplasts was examined after several concentrations of TSA treatments (0, 0.1, 1 and 10 µM). RNP delivery from protoplasts was conducted by conventional polyethylene glycol (PEG) transfection protocols. Interestingly, the indel frequency of the SOC1 gene from TSA treatments was about 3.3 to 3.8 times higher than DMSO treatment in lettuce protoplasts. The TSA-mediated increase of indel frequency of the SOC1 gene in lettuce protoplasts occurred in a concentration-dependent manner, although there was not much difference. Similar to lettuce, TSA also increased the indel frequency by 1.5 to 1.8 times in a concentration-dependent manner during PDS genome editing using tobacco protoplasts. The MNase test clearly showed that chromatin accessibility with TSA treatments was higher than that of DMSO treatment. Additionally, TSA treatment significantly increased the level of histone H3 and H4 acetylation from lettuce protoplasts. The qRT-PCR analysis showed that expression of cell division-related genes (LsCYCD1-1, LsCYCD3-2, LsCYCD6-1, and LsCYCU4-1) was increased by TSA treatment. These findings could contribute to increasing the efficiency of CRISPR/Cas9-mediated genome editing. Furthermore, this could be applied for the development of useful genome-edited crops using the CRISPR/Cas9 system with plant protoplasts.


Assuntos
Sistemas CRISPR-Cas , Edição de Genes/métodos , Ácidos Hidroxâmicos/farmacologia , Alface/metabolismo , Proteínas de Plantas/metabolismo , Protoplastos/metabolismo , Tabaco/metabolismo , Divisão Celular , Genoma de Planta , Alface/efeitos dos fármacos , Alface/genética , Alface/crescimento & desenvolvimento , Células Vegetais , Proteínas de Plantas/antagonistas & inibidores , Proteínas de Plantas/genética , Inibidores da Síntese de Proteínas/farmacologia , Protoplastos/efeitos dos fármacos , Tabaco/efeitos dos fármacos , Tabaco/genética , Tabaco/crescimento & desenvolvimento
5.
Anim Cells Syst (Seoul) ; 25(3): 171-181, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34262660

RESUMO

To identify the specific region of eCG involved in FSH-like activity, the following mutant expression vectors were constructed targeting the amino acid residues 102-104 of the eCG ß-subunit: single mutants, eCGßV102G/α, eCGßF103P/α, and eCGßR104K/α; double mutants, eCGßV102G;F103P/α, eCGßV102G;R104K/α, and eCGßF103P;R104K/α; triple mutant, eCGßV102G;F103P;R104K/α. The LH-like and FSH-like activities of eCG mutants were examined in CHO-K1 cells expressing rat LH/CG receptor and rat FSH receptor. The levels of eCGßV102G/α, eCGßR104K/α, and eCGßV102G;R104K/α in the culture supernatant were markedly lower than those of eCGß/α-wt. The other mutants and rec-eCGß/α-wt were efficiently secreted into the culture supernatant. The LH-like activities of eCGV104G/α, eCGßV102G;R104K/α, and eCGßF103P;R104K/α were approximately 61%, 52%, and 54%, respectively, of those of eCG-wt. The Rmax values of the mutants were 58.9%-78.8% those of eCG-wt with eCGßR104K/α exhibiting the lowest value. The FSH-like activities of single mutants were only 16%-20% of those of eCG-wt. Additionally, the FSH-like activity of double mutants was less than 10% of that of eCG-wt. In particular, the FSH-like activities of ßV102G;R104K/α and ßF103P;R104K/α were 2.5-2.9% of that of eCG-wt. These results suggest that the amino acid residues 102-104 of the eCG ß-subunit are dispensable and that the residue 104 of the eCG ß-subunit plays a pivotal role in signal transduction through the rat FSH receptor. Thus, these mutants may aid future studies on eCG interactions with mammalian FSH receptors in vitro and in vivo.

6.
Biochem Biophys Res Commun ; 563: 40-46, 2021 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-34058473

RESUMO

Standard chemotherapy for ovarian cancers is often abrogated by drug resistance. Specifically, resistance to cisplatin is a major clinical obstacle to successful treatment of ovarian cancers. The aim of this study was to develop a therapeutic strategy using natural killer (NK) cells to treat cisplatin-resistant ovarian cancers. First, we compared the responses of ovarian cancer cell line A2780 and its cisplatin-resistant counterpart, A2780cis, to treatment with cisplatin plus NK92MI cells. Although combined treatment induces apoptosis of ovarian cancer cells via ROS-dependent and -independent mechanisms, A2780cis were resistant to NK92MI cell-mediated cytotoxicity. We found that A2780cis cells showed markedly higher expression of immune checkpoint protein, PD-L1, than the parental cells. Although pretreatment of A2780cis cells with cisplatin stimulated further expression of PD-L1, it also increased expression of ULBP ligands, which are activating receptors on NK92MI cells, both in vitro and in vivo. These findings suggest that combined use of cisplatin plus NK cell-mediated immunotherapy could overcome immunoresistance of chemoresistant ovarian cancers.

7.
Ultrasound Med Biol ; 47(1): 95-103, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33097314

RESUMO

This study aimed to investigate the effect of calcifications on shear-wave elastography in evaluating breast lesions. We retrospectively reviewed ultrasound images of 673 breast lesions and compared the elasticity between lesions with and without calcifications in three subgroups: benign lesions, in situ carcinomas and invasive carcinomas. Breast lesions were confirmed histologically (n = 401) or by follow-up images for more than 2 y (n = 272). Calcifications were present in 25.3% (170/673) of the lesions. The Emean values with and without calcifications, respectively, were as follows: 62.8 and 29.8 kPa in benign lesions (p = 0.000), 114.6 and 52.8 kPa in in situ carcinomas (p = 0.037) and 171.9 and 146.4 kPa in invasive carcinomas (p = 0.018). The presence of calcifications significantly increased the Emean of breast lesions. Shear-wave elastography should be carefully interpreted in benign lesions with calcifications and in situ carcinomas without calcifications.


Assuntos
Doenças Mamárias/complicações , Doenças Mamárias/diagnóstico por imagem , Neoplasias da Mama/complicações , Neoplasias da Mama/diagnóstico por imagem , Calcinose/diagnóstico por imagem , Técnicas de Imagem por Elasticidade , Ultrassonografia Mamária/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Pessoa de Meia-Idade , Estudos Retrospectivos , Adulto Jovem
8.
ACS Appl Mater Interfaces ; 12(51): 56731-56740, 2020 Dec 23.
Artigo em Inglês | MEDLINE | ID: mdl-33290037

RESUMO

Natural killer (NK) cells have been recognized as a next-generation therapy for cancer as they are less likely to trigger adverse events (e.g., cytokine storm or graft-versus-host disease) than T cell-based therapeutics. Although NK cell activation strategies through genetic engineering and cytokine treatment have been actively studied for successful cancer treatment, the approaches are inefficient, expensive, and involve complex processing. Here, we developed a facile and efficient method of activating NK cells using cationic nanoparticles (cNPs). The cytotoxic activity of cNP-treated primary NK and NK-92MI cells against triple-negative breast cancer cells was over 2-fold higher than that of control NK cells in vitro. Molecular biological analyses confirmed that cNPs altered the expression of CCR4 and CXCR4 of NK cells that function as chemokine receptors. In vitro live cell imaging showed that the NK cells treated with cNPs were better than control NK cells at interacting with cancer cells. Consistent with these in vitro results, cNP-treated NK cells effectively inhibited tumor growth in an in vivo tumor animal model of triple-negative breast cancer. Additionally, NK cells treated with cNPs were tracked effectively in vivo by magnetic resonance imaging. Thus, cNP-mediated activation of NK cells has great potential as an NK cell-based cancer immunotherapy. Most of all, activating NK cells using cNPs has a great advantage over conventional methods in that immune cells can be activated by a one-step facile process with exogenously charged nanomaterials, without the need for genetic engineering or cytokine treatment.


Assuntos
Imunoterapia/métodos , Células Matadoras Naturais/imunologia , Ativação Linfocitária/efeitos dos fármacos , Nanopartículas/química , Neoplasias/terapia , Polietilenoimina/farmacologia , Animais , Linhagem Celular Tumoral , Feminino , Humanos , Fatores Imunológicos/química , Fatores Imunológicos/farmacologia , Indóis/química , Células Matadoras Naturais/efeitos dos fármacos , Camundongos Nus , Polietilenoimina/química , Polímeros/química , Ensaios Antitumorais Modelo de Xenoenxerto
9.
Plants (Basel) ; 9(12)2020 Dec 07.
Artigo em Inglês | MEDLINE | ID: mdl-33297321

RESUMO

We aimed to develop a novel technology capable of rapidly selecting mutant plant cell lines. Salt resistance was chosen as a rapid selection trait that is easily applicable to protoplast-derived cell colonies. Mesophyll protoplasts were cultured in a medium supplemented with 0, 50, 100, 150, 200, 250, and 300 mM NaCl. At NaCl concentrations ≥ 100 mM, cell colony formation was strongly inhibited after 4 weeks of culture. Tobacco protoplasts irradiated with 0, 50, 100, 200, and 400 Gy were then cultured to investigate the effects of radiation intensity on cell division. The optimal radiation intensity was 50 Gy. To develop salt-resistant tobacco mutant plants, protoplasts irradiated with 50 Gy were cultured in a medium containing 100 mM NaCl. The efficiency of cell colony formation from these protoplasts was approximately 0.002%. A salt-resistant mutant callus was selected and proliferated in the same medium and then transferred to a shoot inducing medium for adventitious shoot formation. The obtained shoots were then cultured in a medium supplemented with 200 mM NaCl and developed into normal plantlets. This rapid selection technology for generating salt-resistant tobacco mutants will be useful for the development of crop varieties resistant to environmental stresses.

10.
Int J Mol Sci ; 21(22)2020 Nov 10.
Artigo em Inglês | MEDLINE | ID: mdl-33182800

RESUMO

Histone acetylation plays an important role in plant growth and development. Here, we investigated the effect of sodium butyrate (NaB), a histone deacetylase inhibitor, on adventitious shoot formation from protoplast-derived calli and cotyledon explants of tobacco (Nicotiana benthamiana) and tomato (Solanum lycopersicum). The frequency of adventitious shoot formation from protoplast-derived calli was higher in shoot induction medium (SIM) containing NaB than in the control. However, the frequency of adventitious shoot formation from cotyledon explants of tobacco under the 0.1 mM NaB treatment was similar to that in the control, but it decreased with increasing NaB concentration. Unlike in tobacco, NaB decreased adventitious shoot formation in tomato explants in a concentration-dependent manner, but it did not have any effect on adventitious shoot formation in calli. NaB inhibited or delayed the expression of D-type cyclin (CYCD3-1) and shoot-regeneration regulatory gene WUSCHEL (WUS) in cotyledon explants of tobacco and tomato. However, compared to that in control SIM, the expression of WUS was promoted more rapidly in tobacco calli cultured in NaB-containing SIM, but the expression of CYCD3-1 was inhibited. In conclusion, the effect of NaB on adventitious shoot formation and expression of CYCD3-1 and WUS genes depended on the plant species and whether the effects were tested on explants or protoplast-derived calli.


Assuntos
Ácido Butírico/farmacologia , Lycopersicon esculentum/efeitos dos fármacos , Lycopersicon esculentum/crescimento & desenvolvimento , Tabaco/efeitos dos fármacos , Tabaco/crescimento & desenvolvimento , Cotilédone/efeitos dos fármacos , Cotilédone/genética , Cotilédone/crescimento & desenvolvimento , Ciclina D/genética , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Genes de Plantas/efeitos dos fármacos , Inibidores de Histona Desacetilases/farmacologia , Lycopersicon esculentum/genética , Proteínas de Plantas/genética , Brotos de Planta/efeitos dos fármacos , Brotos de Planta/genética , Brotos de Planta/crescimento & desenvolvimento , Especificidade da Espécie , Tabaco/genética
11.
J Nanosci Nanotechnol ; 20(12): 7724-7729, 2020 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-32711649

RESUMO

Despite the fact that stability is a critical issue affecting halide perovskite after the materials have been developed, these materials continue to be studied due to their outstanding optoelectronic characteristics such as narrow emission band width, high PLQY. Many methods are suggested and improved, but the limitations for the display and lighting applications are still remaining. Here, we propose the fabrication of stable cesium lead tri-halide (CsPbX3; X= Cl, Br, I) perovskite films using photocurable polyurethane material, norland optical adhesive 63 (NOA 63), to generate white LEDs by placing films on the InGaN 450 nm blue chip. Comparing with the conventional perovskites, fabricated films well maintained the luminescence properties such as full widths at half maximum (FWHM) of 18 nm and 31 nm for green and red films, respectively. For the stability issue, pristine perovskite without encapsulation is decomposed immediately at high humidity and temperature, but NOA 63 encapsulated perovskite maintained a PL emission property of 60% after four hours in artificial atmosphere. The CIE color triangle reached ~119% of the NTSC standard, exhibiting high color purity. From the results, we confirm that the NOA 63 encapsulated halide perovskites are beneficial when applied in optoelectronic applications due to their improved stability and maintained characteristics.

12.
ACS Nano ; 14(8): 9796-9806, 2020 08 25.
Artigo em Inglês | MEDLINE | ID: mdl-32628447

RESUMO

Sensory adaptation is an essential part of biological neural systems for sustaining human life. Using the light-induced halide phase segregation of CsPb(Br1-xIx)3 perovskite, we introduce neuromorphic phototransistors that emulate human sensory adaptation. The phototransistor based on a hybrid structure of perovskite and transition-metal dichalcogenide (TMD) emulates the sensory adaptation in response to a continuous light stimulus, similar to the neural system. The underlying mechanism for the sensory adaptation is the halide segregation of the mixed halide perovskites. The phase separation under visible-light illumination leads to the segregation of I and Br into separate iodide- and bromide-rich domains, significantly changing the photocurrent in the phototransistors. The devices are reversible upon the removal of the light stimulation, resulting in near-complete recovery of the photosensitivity before the phase segregation (sensitivity recovery of 96.65% for 5 min rest time). The proposed phototransistor based on the perovskite-TMD hybrid structure can be applied to other neuromorphic devices such as neuromorphic photonic devices, intelligent sensors, and selective light-detecting image sensors.


Assuntos
Iodetos , Molibdênio , Compostos de Cálcio , Humanos , Óxidos , Titânio
13.
Int J Mol Sci ; 21(15)2020 Jul 26.
Artigo em Inglês | MEDLINE | ID: mdl-32722633

RESUMO

Enhancing the competence for plant regeneration in tissue culture studies is an important issue not only for efficient genetic transformation of commercial crops but also for the reproducibility of scientific reports. In this study, we investigated optimization of several tissue culture conditions including plant growth regulators, types and ages of explants, culture densities, and plant position in order to improve the competence of adventitious shoot formation of the tomato (Solanum lycopersicum cv. Micro-Tom). In addition, we examined the differential expression of D-type cyclin (CYCD3-1) and several shoot regeneration regulatory genes from hypocotyl and cotyledon explants of tomato during shoot organogenesis. A treatment of 1 mg L-1 Zeatin and 0.1 mg L-1 Indole-3-acetic acid (IAA) in Murashige and Skoog (MS) medium containing 3% sucrose was optimal for adventitious shoot formation from hypocotyl and cotyledon explants. The younger explants exhibited more shoot formation regardless of explant types. Additionally, those closest to the shoot apical meristem produced more shoots compared to the other regions in the hypocotyl and the cotyledon explants. Gene expression of CYCD3-1, SHOOT MERISTEMLESS (STM), and cytokinin dependent WUSCHEL (WUS) was significantly higher in younger explants than in older ones. Furthermore, an increase in CYCD3-1, STM, and WUS expression was evident at the distal part of hypocotyls and the proximal part of cotyledons compared to other regions. These differential gene expression profiles exhibited good agreement with the results of shoot formation obtained from diverse explants of tomato. These results suggest that temporal and spatial gene expression of shoot regeneration regulatory genes plays an important role in enhancing the competence and the reproducibility of adventitious shoot formation from tomato explants.


Assuntos
Cotilédone/metabolismo , Regulação da Expressão Gênica de Plantas , Hipocótilo/metabolismo , Lycopersicon esculentum/metabolismo , Proteínas de Plantas/biossíntese
14.
Behav Med ; : 1-8, 2020 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-32701418

RESUMO

Four common health risk behaviors have the greatest impact on all-cause mortality risk, but studies are needed with larger samples and the appropriate age range for cigarette smokers. We examined the impact of smoking in the context of multiple health behaviors on all-cause mortality using a nationally representative sample of adults aged 30 and older in the United States. National Health Interview Survey data from 1997 to 2005 were linked to the National Death Index with a follow-up to December 2015. The primary dependent variable was all-cause mortality, and the primary predictors were smoking, heavy drinking, physical inactivity, and unhealthy weight (underweight or obesity). The sample contained 189,087 individuals (≥ age 30; population estimate = 140.7 million). Our primary statistical analysis tool involved fitting Cox proportional hazards models. Our findings demonstrated that smoking led to the highest mortality risk among the four risk behaviors examined, but more than half of smokers engaged in at least one additional health risk behavior. Smokers who engaged in multiple health behaviors experienced higher increased mortality risks: smoking combined with one other health risk behavior increased mortality risk by 32% and by 82% when combined with two behaviors. Engaging in all four risk behaviors more than doubled the mortality risk of smokers. Smoking cessation interventions that address multiple risk behaviors-physical inactivity, heavy drinking, and unhealthy weight-will likely prevent premature death better than interventions that address only smoking.

15.
Cancers (Basel) ; 12(5)2020 May 13.
Artigo em Inglês | MEDLINE | ID: mdl-32414208

RESUMO

Triple-negative breast cancer (TNBC) refers to breast cancer that does not have receptors for estrogen, progesterone, and HER2 protein. TNBC accounts for 10-20% of all cases of breast cancers and is characterized by its metastatic aggressiveness, poor prognosis, and limited treatment options. Here, we show that the metastatic nature of TNBC is critically regulated by a functional link between miR-200a and the transcription factor ELK3. We found that the expression levels of miR-200a and the ELK3 mRNA were negatively correlated in the luminal and TNBC subtypes of breast cancer cells. In vitro experiments revealed that miR-200a directly targets the 3' untranslated region (UTR) of the ELK3 mRNA to destabilize the transcripts. Furthermore, ectopic expression of miR-200a impaired the migration and invasion of TNBC cells by reducing the expression level of the ELK3 mRNA. In in vivo studies, transfection of MDA-MB 231 cells (a claudin-low TNBC cell type) with exogenous miR-200a reduced their extravasation into the lung during 48 h after tail vein injection, and co-transfection of the cells with an expression plasmid harboring ELK3 that lacked an intact 3'UTR recovered their extravasation ability. Overall, our findings provide evidences that miR-200a and ELK3 is functionally linked to regulate invasive characteristics of breast cancers.

16.
Public Health Nurs ; 37(2): 161-168, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31724240

RESUMO

OBJECTIVES: To examine smoking behavior and social contexts related to smoking among dual-smoker couples. DESIGN: Cross-sectional online survey study. SAMPLE: A convenience sample of 183 dual-smoker couples. MEASUREMENTS: Investigator-developed survey on smoking and related social contexts. RESULTS: Participants smoked 16.0 cigarettes daily for 14.2 years; 48.4% shared more than half of their smoking time with their spouse. More than half made quit attempts in the past year individually (M = 5.3) and jointly (M = 2.5). Couples sharing more smoking time were more likely to be motivated to quit (p = .002), make quit attempts (p < .0001), and be interested in cessation interventions (p = .002); but less likely to implement home smoking bans (p < .001). Among those who reported quit attempts, 41% quit by themselves and 15.3% sought professional assistance. Most common reasons for relapse were chronic stress and crisis, 63.6%, however, were interested in smoking cessation services, preferably technology-based interventions. CONCLUSIONS: We found smoking interdependence within dual-smoker couples. Despite high levels of motivation to quit, most did not utilize professional help, leading to low successful quit rates. Technology-based smoking cessation interventions incorporating spousal support and addressing stress/crisis may best assist dual-smoker couples.


Assuntos
Fumar/psicologia , Meio Social , Cônjuges/psicologia , Adulto , Estudos Transversais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Motivação , Abandono do Hábito de Fumar/psicologia , Abandono do Hábito de Fumar/estatística & dados numéricos , Cônjuges/estatística & dados numéricos , Inquéritos e Questionários , Adulto Jovem
17.
Sci Rep ; 9(1): 16354, 2019 11 08.
Artigo em Inglês | MEDLINE | ID: mdl-31704990

RESUMO

Biocontrol offers a promising alternative to synthetic fungicides for the control of a variety of pre- and post-harvest diseases of crops. Black rot, which is caused by the pathogenic fungus Ceratocytis fimbriata, is the most destructive post-harvest disease of sweet potato, but little is currently known about potential biocontrol agents for this fungus. Here, we isolated several microorganisms from the tuberous roots and shoots of field-grown sweet potato plants, and analyzed their ribosomal RNA gene sequences. The microorganisms belonging to the genus Pantoea made up a major portion of the microbes residing within the sweet potato plants, and fluorescence microscopy showed these microbes colonized the intercellular spaces of the vascular tissue in the sweet potato stems. Four P. dispersa strains strongly inhibited C. fimbriata mycelium growth and spore germination, and altered the morphology of the fungal hyphae. The detection of dead C. fimbriata cells using Evans blue staining suggested that these P. dispersa strains have fungicidal rather than fungistatic activity. Furthermore, P. dispersa strains significantly inhibited C. fimbriata growth on the leaves and tuberous roots of a susceptible sweet potato cultivar ("Yulmi"). These findings suggest that P. dispersa strains could inhibit black rot in sweet potato plants, highlighting their potential as biocontrol agents.


Assuntos
Ascomicetos/crescimento & desenvolvimento , Ipomoea batatas/imunologia , Pantoea/fisiologia , Controle Biológico de Vetores , Doenças das Plantas/prevenção & controle , Folhas de Planta/imunologia , Raízes de Plantas/imunologia , Resistência à Doença , Ipomoea batatas/microbiologia , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Folhas de Planta/microbiologia , Raízes de Plantas/microbiologia
18.
Sci Rep ; 9(1): 15397, 2019 10 28.
Artigo em Inglês | MEDLINE | ID: mdl-31659261

RESUMO

Scrub typhus is an acute febrile illness caused by obligate intracellular organism Orientia tsutsugamushi. While there have been many reports on the evaluation of disease activity and infectious diseases using F-18 fluorodeoxyglucose (FDG) positron emission tomography/computed tomography (PET/CT), the clinical value of FDG PET/CT in scrub typhus has not been fully investigated. We enrolled 17 patients who were 18 years of age or older and clinically suspected of having scrub typhus with eschar. Clinical assessments, blood samples, and FDG PET/CT images were obtained at enrolment and again after 3 weeks. The median age of the patients was 65 years; 9 (52.9%) patients were male. On initial FDG PET/CT, the eschars showed markedly increased FDG uptake on PET imaging that improved after treatment. Generalized lymphadenopathy and splenomegaly with high FDG uptake were observed in all patients. On follow-up FDG PET/CT after appropriate therapy, FDG uptake and sizes of eschar, lymph nodes, and spleen were markedly decreased. As far as we are aware, this is the first investigation with multiple patients of FDG PET/CT in scrub typhus and the demonstration of clinical utility. FDG PET/CT imaging of scrub typhus could provide useful information about the clinical features before and after antibiotic treatment.


Assuntos
Antibacterianos/uso terapêutico , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada , Tifo por Ácaros/diagnóstico por imagem , Idoso , Antibacterianos/administração & dosagem , Feminino , Fluordesoxiglucose F18 , Humanos , Linfonodos/diagnóstico por imagem , Masculino , Pessoa de Meia-Idade , Compostos Radiofarmacêuticos , Tifo por Ácaros/tratamento farmacológico , Baço/diagnóstico por imagem
19.
Biomaterials ; 221: 119418, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31419655

RESUMO

Recently, natural killer (NK)-based immunotherapy has attracted attention as a next-generation cell-based cancer treatment strategy due to its mild side effects and excellent therapeutic efficacy. Here, we describe multifunctional nanoparticles (MF-NPs) capable of genetically manipulating NK cells and tracking them in vivo through non-invasive magnetic resonance (MR) and fluorescence optical imaging. The MF-NPs were synthesized with a core-shell structure by conjugation of a cationic polymer labeled with a near-infrared (NIR) fluorescent molecule, with the aid of a polydopamine (PDA) coating layer. When administered to NKs, the MF-NPs exhibited excellent cytocompatibility, efficiently delivered genetic materials into the immune cells, and induced target protein expression. In particular, the MF-NPs could induce the expression of EGFR targeting chimeric antigen receptors (EGFR-CARs) on the NK cell surface, which improved the cells' anti-cancer cytotoxic effect both in vitro and in vivo. Finally, when NK cells labeled with MF-NPs were injected into live mice, MF-NP-labeled NK cells could be successfully imaged using fluorescence and MR imaging devices. Our findings indicate that MF-NPs have great potential for application of NK cells, as well as other types of cell therapies involving genetic engineering and in vivo monitoring of cell trafficking.


Assuntos
Células Matadoras Naturais/citologia , Nanopartículas Multifuncionais/química , Linhagem Celular Tumoral , Sobrevivência Celular/fisiologia , Citometria de Fluxo , Engenharia Genética/métodos , Humanos , Imunoterapia Adotiva/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa
20.
Radiology ; 292(1): 37-48, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31038406

RESUMO

Background The relationship between method of breast cancer screening (mammography or US) and survival outcome in patients with screening-detected ductal carcinoma in situ (DCIS) has not been determined. Purpose To investigate whether different methods of breast cancer screening are associated with different survival outcomes in patients with screening-detected DCIS and to evaluate clinical-pathologic and imaging factors associated with second breast cancer. Materials and Methods We retrospectively identified women who underwent surgery to treat DCIS initially detected with screening mammography or US between July 2004 and December 2011 in a single institution. Overall survival (OS) and disease-free survival (DFS) were assessed. Factors associated with second breast cancer (invasive carcinoma or DCIS) were found with multivariable Cox proportional hazards regression analysis. Subgroups were analyzed according to screening method. Results A total of 814 women (median age, 47 years; age range, 25-81 years) were included; 627 underwent treatment for screening mammography-detected DCIS (mammography-detected group), and 187 underwent treatment for screening US-detected DCIS (US-detected group). During follow-up (median, 7 years; interquartile range, 5-8 years), 26 ipsilateral and 26 contralateral second breast cancers (6.4%, 52 of 814) were found, with 44 in the mammography-detected group and eight in the US-detected group. The overall 5-year OS and DFS rates were 100% and 95.3%, respectively. DFS rates did not differ according to screening method (P = .21, 5-year DFS rates were 94.9% in the mammography-detected group and 96.5% in the US-detected group). In the mammography-detected group, higher nuclear grade (intermediate grade: hazard ratio [HR], 5.7; 95% confidence interval [CI]: 1.3, 24.3; P = 0.02) (high grade: HR, 8.0; 95% CI: 1.9, 34.2; P = .01) and dense breast (HR, 3.5; 95% CI: 1.1, 11.4; P = 0.04) were associated with second breast cancer. In the US-detected group, human epidermal growth factor receptor 2 positivity was associated with second breast cancer (HR, 9.2; 95% CI: 2.2, 38.5; P = .002). Conclusion Disease-free survival of patients who underwent treatment for screening-detected ductal carcinoma in situ (DCIS) did not differ according to screening detection method. In patients with screening mammography-detected DCIS, higher nuclear grade and dense breast were associated with second breast cancer, and in patients with screening US-detected DCIS, human epidermal growth factor 2 positivity was associated with second breast cancer. © RSNA, 2019 Online supplemental material is available for this article. See also the editorial by Fowler in this issue.


Assuntos
Neoplasias da Mama/diagnóstico por imagem , Carcinoma Intraductal não Infiltrante/diagnóstico por imagem , Mamografia/métodos , Segunda Neoplasia Primária/diagnóstico , Ultrassonografia Mamária/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Mama/diagnóstico por imagem , Feminino , Seguimentos , Humanos , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Estudos Retrospectivos , Taxa de Sobrevida
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