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An ABCDmatrix representation in space and time of a mode-locked laser is presented. The conventional stability concept of Gaussian beams in laser cavities implies, instead of stability, an equilibrium condition, which can be indifferent, stable, or unstable. It is further shown that when beam and pulse evolution in the laser are intertwined, stable mode locking can be the result of evolution of a noise spike in an otherwise unstable (in the conventional sense) cavity.
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Plant volatile organic compounds (VOCs) are an important link that mediates chemical communication between plants and plants, plants and insects, and plants and natural enemies of insect pests. In this study, we tested the response in the selective behavior of western flower thrips, Frankliniella occidentalis, to the VOCs of kidney bean, Phaseolus vulgaris L., to explore their "attraction" or "repellent" effects regarding their application in integrated pest management (i.e., IPM). The results indicated that 12.7 µL/mL (E, E, E, E)-squalene, 3.2 µL/mL dioctyl phthalate, and 82.2 µL/mL ethyl benzene had a significantly attractive effect on the selective behavior of F. occidentalis, while 10.7 µL/mL and 21.4 µL/mL 2,6-ditert-butyl-4-methyl phenol had a significantly repulsive effect on the selective behavior of F. occidentalis, showing that F. occidentalis responds differently to specific concentrations of VOCs from P. vulgaris plant emissions. Interestingly, the three compounds with the specific above concentrations, after being mixed in pairs, significantly attracted F. occidentalis compared to the control treatment; however, the mixture with the three above compounds had no significant different effect on F. occidentalis compared to the control treatment. It can be seen that the effect with the mixtures of three kinds of VOCs had the same function and may not get better. Simultaneously, the reasons for this result from the transcription levels of odorant-binding protein genes (OBPs) were determined. There were differences in the types and transcription levels of OBPs, which played a major role in the host selection behavior of F. occidentalis under the mixed treatment of different VOCs. It is presumed that there are specific VOCs from P. vulgaris plants that have a good repellent or attracting effect on the selective behavior of F. occidentalis, which can be used for the development of plant-derived insect attractants and repellents to serve as IPM in fields. But attention should be paid to the antagonism between plant-derived preparations and VOCs produced by plants themselves after application.
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Repelentes de Insectos , Phaseolus , Thysanoptera , Compuestos Orgánicos Volátiles , Animales , Compuestos Orgánicos Volátiles/farmacología , Verduras , FloresRESUMEN
Previous studies have identified GAPDH as a promising target for treating cancer and modulating immunity because its inhibition reduces glycolysis in cells (cancer cells and immune cells) with the Warburg effect, a modified form of cellular metabolism found in cancer cells. However, the quantitative relationship between GAPDH and the aerobic glycolysis remains unknown. Here, using siRNA-mediated knockdown of GAPDH expression and iodoacetate-dependent inhibition of enzyme activity, we examined the quantitative relationship between GAPDH activity and glycolysis rate. We found that glycolytic rates were unaffected by the reduction of GAPDH activity down to 19% ± 4.8% relative to untreated controls. However, further reduction of GAPDH activity below this level caused proportional reductions in the glycolysis rate. GAPDH knockdown or inhibition also simultaneously increased the concentration of glyceraldehyde 3-phosphate (GA3P, the substrate of GAPDH). This increased GA3P concentration countered the effect of GAPDH knockdown or inhibition and stabilized the glycolysis rate by promoting GAPDH activity. Mechanistically, the intracellular GA3P concentration is controlled by the Gibbs free energy of the reactions upstream of GAPDH. The thermodynamic state of the reactions along the glycolysis pathway was only affected when GAPDH activity was reduced below 19% ± 4.8%. Doing so moved the reactions catalyzed by GAPDH + PGK1 (phosphoglycerate kinase 1, the enzyme immediate downstream of GAPDH) away from the near-equilibrium state, revealing an important biochemical basis to interpret the rate control of glycolysis by GAPDH. Collectively, we resolved the numerical relationship between GAPDH and glycolysis in cancer cells with the Warburg effect and interpreted the underlying mechanism.
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Gliceraldehído-3-Fosfato Deshidrogenasa (Fosforilante)/fisiología , Glucólisis/fisiología , Neoplasias/metabolismo , Línea Celular Tumoral , Glucosa/metabolismo , Gliceraldehído-3-Fosfato Deshidrogenasa (Fosforilante)/genética , Gliceraldehído-3-Fosfato Deshidrogenasa (Fosforilante)/metabolismo , Gliceraldehído-3-Fosfato Deshidrogenasas/metabolismo , Humanos , Oxidación-Reducción , ARN Interferente Pequeño/genética , Efecto Warburg en OncologíaRESUMEN
This study addresses any sensor based on measuring a physical quantity through the phase of a probing beam. This includes sensing of rotation, acceleration, index change, displacement, fields While most phase measurements are made by detecting an amplitude change in interfering beams, we detect instead a phase change through a relative frequency shift of two correlated frequency combs. This paper explores the limit sensitivity that this method can achieve, when the combs are generated in an Optical Parametric Oscillator (OPO), pumped synchronously by a train of femtosecond pulses separated by half the OPO cavity round-trip time. It is shown that a phase difference as small as 0.4 nanoradians can be resolved between the two pulses circulating in the cavity. This phase difference is one order of magnitude better than the previous record. The root-mean-square deviation of the measured phase over measuring time is close to the standard quantum limit (phase-photon number uncertainty product of 0.66). Innovations that made such improved performances possible include a more stable OPO cavity design; a stabilization system with a novel purely electronic locking of the OPO cavity length relative to that of the pump laser; a shorter pump laser cavity; and a square pulse generator for driving a 0.5 mm pathlength lithium niobate phase modulator. Future data acquisition improvements are suggested that will bring the phase sensitivity exactly to the standard quantum limit, and beyond the quantum limit by squeezing.
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Phosphoglycerate kinase 1 (PGK1) plays important roles in glycolysis, yet its forward reaction kinetics are unknown, and its role especially in regulating cancer cell glycolysis is unclear. Here, we developed an enzyme assay to measure the kinetic parameters of the PGK1-catalyzed forward reaction. The Km values for 1,3-bisphosphoglyceric acid (1,3-BPG, the forward reaction substrate) were 4.36 µm (yeast PGK1) and 6.86 µm (human PKG1). The Km values for 3-phosphoglycerate (3-PG, the reverse reaction substrate and a serine precursor) were 146 µm (yeast PGK1) and 186 µm (human PGK1). The Vmax of the forward reaction was about 3.5- and 5.8-fold higher than that of the reverse reaction for the human and yeast enzymes, respectively. Consistently, the intracellular steady-state concentrations of 3-PG were between 180 and 550 µm in cancer cells, providing a basis for glycolysis to shuttle 3-PG to the serine synthesis pathway. Using siRNA-mediated PGK1-specific knockdown in five cancer cell lines derived from different tissues, along with titration of PGK1 in a cell-free glycolysis system, we found that the perturbation of PGK1 had no effect or only marginal effects on the glucose consumption and lactate generation. The PGK1 knockdown increased the concentrations of fructose 1,6-bisphosphate, dihydroxyacetone phosphate, glyceraldehyde 3-phosphate, and 1,3-BPG in nearly equal proportions, controlled by the kinetic and thermodynamic states of glycolysis. We conclude that perturbation of PGK1 in cancer cells insignificantly affects the conversion of glucose to lactate in glycolysis.
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Glucólisis , Proteínas de Neoplasias , Neoplasias , Fosfoglicerato Quinasa , Células A549 , Ácidos Difosfoglicéricos/química , Ácidos Difosfoglicéricos/metabolismo , Glucosa/química , Glucosa/metabolismo , Ácidos Glicéricos/química , Ácidos Glicéricos/metabolismo , Células HeLa , Humanos , Cinética , Ácido Láctico/química , Ácido Láctico/metabolismo , Proteínas de Neoplasias/química , Proteínas de Neoplasias/metabolismo , Neoplasias/química , Neoplasias/metabolismo , Fosfoglicerato Quinasa/química , Fosfoglicerato Quinasa/metabolismo , Saccharomyces cerevisiae/química , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/química , Proteínas de Saccharomyces cerevisiae/metabolismoRESUMEN
Background: Intestinal ischemia/reperfusion (I/R) injury is a common tissue-organ damage occurring in surgical practice. This study aims to comprehensively review the collaboration and impact of countries, institutions, authors, subject areas, journals, keywords, and critical literature on intestinal I/R injury from a bibliometric perspective, and to assess the evolution of clustering of knowledge structures and identify hot trends and emerging topics. Methods: Articles and reviews related to intestinal I/R were retrieved through subject search from Web of Science Core Collection. Bibliometric analyses were conducted on Excel 365, CiteSpace, VOSviewer, and Bibliometrix (R-Tool of R-Studio). Results: A total of 1069 articles and reviews were included from 2004 to 2022. The number of articles on intestinal I/R injury gradually plateaued, but the number of citations increased. These publications were mainly from 985 institutions in 46 countries, led by China and the United States. Liu Kx published the most papers, while Chiu Cj had the largest number of co-citations. Analysis of the journals with the most outputs showed that most journals focused on surgical sciences, cell biology, and immunology. Macroscopic sketch and microscopic characterization of the entire knowledge domain were achieved through co-citation analysis. The roles of cell death, exosomes, intestinal flora, and anesthetics in intestinal I/R injury are the current and developing research focuses. The keywords "dexmedetomidine", "proliferation", and "ferroptosis" may also become new trends and focus of future research. Conclusion: This study comprehensively reviews the research on intestinal I/R injury using bibliometric and visualization methods, and will help scholars better understand the dynamic evolution of intestinal I/R injury and provide directions for future research.
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Although glucose, through pentose phosphate pathway (PPP), is the main source to generate NADPH, solid tumors are often deprived of glucose, hence alternative metabolic pathways to maintain NADPH homeostasis in cancer cells are required. Here, we report that lactate and glutamine support NADPH production via isocitrate dehydrogenase 1 (IDH1) and malic enzyme 1 (ME1), respectively, under glucose-deprived conditions. Isotopic tracing demonstrates that lactate participates in the formation of isocitrate. Malate derived from glutamine in mitochondria shuttles to cytosol to produce NADPH. In cells cultured in the absence of glucose, knockout of IDH1 and ME1 decreases NADPH/NADP+ and GSH/GSSG, increases ROS level and facilitates cell necrosis. In 4T1 murine breast tumors, knockout of ME1 retards tumor growth in vivo, with combined ME1/IDH1 knockout more strongly suppressing tumor growth. Our findings reveal two alternative NADPH-producing pathways that cancer cells use to resist glucose starvation, reflecting the metabolic plasticity and flexibility of cancer cells adapting to nutrition stress.
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Glucosa , Neoplasias , Animales , Glutamina , Ácido Láctico , Ratones , NADP/metabolismo , Neoplasias/genética , Vía de Pentosa FosfatoRESUMEN
The effects of industrial storage on the changes of the cell viability and the activities of intracellular alcohol dehydrogenase (ADH) and glucose-6-phosphate dehydrogenase (G6PDH) in brewer's yeast, and the corresponding capacity for the bioconversion of ethyl-3-oxobutanoate (EOB) to ethyl (S)-3-hydroxybutanoate ((S)-EHB), were investigated. The viability of fresh brewer's yeast cells stored in industrial circulating cooling water at 1-2 degrees C showed 4 and 15% drop after the storage of 7 and 15 days, respectively, after which cells died rapidly. The pretreatment of the stored brewer's yeast cells by washing and screening significantly enhanced cell viability during industrial storage. The intracellular levels of ADH and G6PDH after permeabilization of these stored cells with cetyltrimetylammonium bromide (CTAB) were much higher, which showed only slight decrease within 2 weeks during the industrial storage. When the stored cells after the permeabilization treatment was used as the biocatalyst at 90-120 g/L, EOB was converted almost completely into enantiopure (S)-EHB with an enantiomeric excess (ee) more than 99% and a yield of over 96%, by fed-batch bioconversion of 560 mM EOB within 6 h.
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Alcohol Deshidrogenasa/metabolismo , Glucosafosfato Deshidrogenasa/metabolismo , Microbiología Industrial , Preservación Biológica/métodos , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/enzimología , Viabilidad MicrobianaRESUMEN
U-shaped sacral fractures are rare and often difficult to diagnose primarily due to the difficulty in obtaining adequate imaging and the severe associated injuries. These fractures are highly unstable and frequently cause neurological deficits. The majority of surgeons have limited experience in management of U-shaped sacral fractures. No standard treatment protocol for U-shaped sacral fractures has been available till now. This study aimed to examine the management of U-shaped sacral fractures and the early outcomes. Clinical data of 15 consecutive patients with U-shaped sacral fracture who were admitted to our trauma center between 2009 and 2014 were retrospectively analyzed. Demographics, fracture classification, mechanism of injury and operative treatment and deformity angle were assessed. All the patients were treated with lumbopelvic fixation or (and) sacral decompression. EQ-5d score was applied to evaluate the patients' quality of life. Of the 15 consecutive patients with U-shaped sacral fracture, the mean age was 28.8 years (range: 15-55 years) at the time of injury. There were 6 females and 9 males. The mean followup time was 22.7 months (range: 9 47 months) and mean full weight-bearing time was 9.9 weeks (range: 8-14 weeks). Ten patients received lumbopelvic fixation and sacral decompression, one lombosacral fixation, and 4 merely sacral decompression due to delayed diagnosis or surgery. The post-operation deformity angle (mean 27.87°, and range: 8°-90°) of the sacrum was smaller than that pre-operation (mean 35.67; range: 15-90) with no significance difference noted. At the latest follow-up, all patients obtained neurological recovery with different extents. Visual analogue score (VAS) was reduced from preoperative 7.07 (range: 5-9) to postoperetive 1.93 (range: 1-3). All patients could walk without any aid after treatment. Eight patients were able to care for themselves and undertook some daily activities. Five patients had returned to work foil time. In conclusion, lumbopelvic fixation is an effective method for stabilization of U-shaped sacral fractures with fewer complications developed. Effective reduction and firm fixation are the prerequisite of early mobilization and neurological recovery. Sacral decompression effectively promotes neurological recovery even in patients with old U-shaped sacral fractures.
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Descompresión Quirúrgica/métodos , Fijación Interna de Fracturas/métodos , Región Lumbosacra/cirugía , Complicaciones Posoperatorias/patología , Fracturas de la Columna Vertebral/cirugía , Adolescente , Adulto , Tornillos Óseos/efectos adversos , Descompresión Quirúrgica/efectos adversos , Femenino , Fijación Interna de Fracturas/efectos adversos , Fijación Interna de Fracturas/instrumentación , Humanos , Región Lumbosacra/lesiones , Masculino , Persona de Mediana Edad , Pelvis/cirugía , Complicaciones Posoperatorias/diagnóstico por imagen , Complicaciones Posoperatorias/epidemiologíaRESUMEN
A self-humidifying membrane based on low-cost sulfonated poly (ether ether ketone) (SPEEK) hybrid with sulfated zirconia (SO4(2-)/ZrO2, SZ)-supported platinum catalyst (Pt-SZ catalyst) was investigated for fuel cell applications. The SZ particle, a solid-state superacid with hygroscopic and high proton conductivity properties, was employed as the catalyst support. The SPEEK/Pt-SZ self-humidifying membrane was characterized by TEM and SEM coupled with EDX. FT-IR was conducted to verify the effect of SPEEK/Pt-SZ membrane on catalytic combination of crossover hydrogen and oxygen. To display the advantages of Pt-SZ catalyst as the additive, the IEC, water uptake, proton conductivity, single-cell performance, and areal resistance measurements were compared between the plain SPEEK membrane, SPEEK/Pt-SiO2 membrane, and the SPEEK/Pt-SZ membrane. The SPEEK/Pt-SZ membrane exhibited the highest IEC value, proton conductivity, single-cell performance, and the lowest areal resistance relative to the plain SPEEK and SPEEK/Pt-SiO2 membranes. The SPEEK/Pt-SZ self-humidifying membrane exhibited peak power density of 1.0 W/cm2 under dry operation condition compared with 0.89 W/cm2 and 0.58 W/cm2 of SPEEK/Pt-SiO2 and plain SPEEK membranes, respectively. The incorporation of the catalytic, hygroscopic and proton conductive Pt-SZ catalyst in the SPEEK/Pt-SZ self-humidifying membrane facilitated water balance and proton conduction, and accordingly improved its single cell performance under dry operation. In addition, the enhanced OCV and the decreased areal ohmic resistance confirmed the promotion effect of Pt-SZ catalyst in the self-humidifying membrane on suppressing reactant crossover and the membrane self-humidification.
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A carbon-supported PtRuNi nanocomposite is synthesized via a microwave-irradiated polyol plus annealing synthesis strategy. The catalyst is characterized by transmission electron microscopy, powder X-ray diffraction, energy dispersive spectroscopy, and X-ray photoelectron spectroscopy. The data are discussed with respect to those for the carbon-supported PtRu nanocomposite prepared following the same way. The characterizations show that the inclusion of Ni in the PtRu system has only a small effect on the particle size, the structure, and the compositional homogeneity. CO-stripping voltammetry and measurements on the single proton exchange membrane fuel cells show that the PtRuNi/C catalyst has an improved activity for CO(ads) electro-oxidation. An accelerated durability test on the catalyst exhibits insignificant loss of activity in acidic media. On the basis of the exploration of the structure-activity relationship, a mechanism for the improved performance of the catalyst is proposed. It is suggested that the improved CO-tolerant performance of the PtRuNi/C nanocomposite should be related to the hydrogen spillover on the catalyst surface, the enhanced oxidation of CO(ads) by nickel hydroxides, and the high proton and electronic conductivity of the hydroxides. The nickel hydroxide passivated surface and/or anchoring of metallic nickel in the platinum lattice may contribute to the durability of the catalyst in acid solution.
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An ultrathin poly(tetrafluoroethylene) (PTFE)-reinforced multilayer self-humidifying composite membrane (20 microm, thick) is developed. The membrane is composed of Nafion-impregnated porous PTFE composite as the central layer, and SiO2 supported nanosized Pt particles (Pt-SiO2) imbedded into the Nafion as the two side layers. The proton exchange membrane (PEM) fuel cell employing the self-humidifying membrane (Pt-SiO2/NP) turns out a peak power density of 1.40 W cm(-2) and an open circuit voltage (OCV) of 1.032 V under dry H2/O2 condition. The excellent performance is attributed to the combined result of both the accelerated water back-diffusion in the thin membrane and the adsorbing/releasing water properties of the Pt-SiO2 catalyst in the side layers. Moreover, the inclusion of the hygroscopic Pt-SiO2 catalyst inside the membrane results in an enhanced anode self-humidification capability and also the decreased cathode polarization (accordingly an improved cell OCV). Several techniques, such as transmission electronic microscopy, scanning electronic microscopy, energy dispersive spectroscopy, thermal analysis and electrochemical impedance spectroscopy etc., are employed to characterize the Pt-SiO2/NP membrane. The results are discussed in comparison with the plain Nafion/PTFE membrane (NP). It is established that the reverse net water drag (from the cathode to the anode) across the Pt-SiO2/NP membrane reaches 0.16 H2O/H+. This implies a good hydration of the Pt-SiO2/NP membrane and thus ensures an excellent PEM fuel cell performance under self-humidification operation.
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Suministros de Energía Eléctrica , Membranas Artificiales , Politetrafluoroetileno/química , Diseño de Equipo , Humedad , Hidrógeno/química , Microscopía Electrónica de Rastreo/métodos , Microscopía Electrónica de Transmisión/métodos , Oxígeno/química , Tamaño de la Partícula , Platino (Metal)/química , Sensibilidad y Especificidad , Dióxido de Silicio/química , Agua/químicaRESUMEN
OBJECTIVE: To investigate the effects of fibular head resection in prosthetic replacement for neoplasms of the proximal tibia in limb salvage surgery. METHODS: Between July 1999 and March 2013, 76 patients with neoplasms of the proximal tibia underwent tumor resection, prosthetic replacement, and gastrocnemius medial head flap transfer. Among them, 38 patients underwent fibular head resection (group A) and 38 underwent fibular head preservation (group B). There was no significant difference in gender, age, side, tumor classification and stage, and disease duration between 2 groups (P > 0.05). The complications and the position of the components were observed, and American society for bone tumors scoring system (MSTS93) was used to evaluate the joint function. RESULTS: All patients were followed up 12-150 months (mean, 87 months). Incision infection occurred in 1 patient (2.63%) of group A and 6 patients (15.79%) of group B, showing significant difference (χ2 = 3.934, P = 0.047). Necrosis of gastrocnemius medial head flap was found in 1 patient of group A and 2 patients of group B. Prosthetic loosening and instability of the knee were observed in 4 and 2 cases of group A and in 6 and 4 cases of group B, respectively. In groups A and B, there were 3 and 5 cases of local recurrence, 7 and 6 cases of distant metastasis, and 8 and 7 deaths, respectively. According to MSTS93, the results were excellent in 23 cases, good in 10 cases, fair in 3 cases, and poor in 2 cases, with an excellent and good rate of 86.84% in group A; the results were excellent in 21 cases, good in 11 cases, fair in 3 cases, and poor in 3 cases, with an excellent and good rate of 84.21% in group B; and no significant difference was found in the excellent and good rate between 2 groups (χ2 = 0.106, P = 0.744). CONCLUSION: Fibular head resection in prosthetic replacement for neoplasms of the proximal tibia in limb salvage surgery is beneficial to intra-operative tissue coverage, and it can reduce trauma by skin transplantation and related complications. Good stability and motion of the joint can be obtained after operation.
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Neoplasias Óseas/cirugía , Recuperación del Miembro , Recurrencia Local de Neoplasia/cirugía , Tibia/cirugía , Epífisis , Peroné , Estudios de Seguimiento , Humanos , Rodilla , Articulación de la Rodilla , Músculo Esquelético , Implantación de Prótesis , Colgajos Quirúrgicos , Resultado del TratamientoRESUMEN
The piperidine alkaloid piperine, a major ingredient in black pepper, inhibits the growth and metastasis of cancer cells both in vivo and in vitro, although its mechanism of action is unclear. Furthermore, its anticancer activity against osteosarcoma cells has not been reported. In this study, we show that piperine inhibited the growth of HOS and U2OS cells in dose- and time-dependent manners but had a weaker effect on the growth of normal hFOB cells. Piperine inhibited osteosarcoma cell proliferation by causing G2/M phase cell cycle arrest associated with decreased expression of cyclin B1 and increased phosphorylation of Cyclin-dependent kinase-1(CDK1) and checkpoint kinase 2 (Chk2). In addition, piperine treatment inhibited phosphorylation of Akt and activated phosphorylation of c-Jun N-terminal kinase (c-JNK) and p38 mitogen-activated protein kinase (MAPK) in HOS and U2OS cells. Piperine induced colony formation in these two cell types. We proved that piperine could suppress the metastasis of osteosarcoma cells using scratch migration assays and Transwell chamber tests. Moreover, gelatin zymography showed that piperine inhibited the activity of matrix metalloproteinase (MMP)-2/-9 and increased the expression of tissue inhibitor of metalloproteinase (TIMP)-1/-2. Taken together, our results indicate that piperine inhibits proliferation, by inducing G2/M cell cycle arrest, and the migration and invasion of HOS and U2OS cells, via increased expression of TIMP-1/-2 and down-regulation of MMP-2/-9. These findings support further study of piperine as a promising therapeutic agent in the treatment of osteosarcoma.
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Alcaloides/uso terapéutico , Benzodioxoles/uso terapéutico , Neoplasias Óseas/tratamiento farmacológico , Inhibidores de Crecimiento/uso terapéutico , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Osteosarcoma/tratamiento farmacológico , Piper nigrum/química , Piperidinas/uso terapéutico , Alcamidas Poliinsaturadas/uso terapéutico , Proteína Quinasa CDC2/metabolismo , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular/efectos de los fármacos , Quinasa de Punto de Control 2/metabolismo , Ciclina B1/genética , Ciclina B1/metabolismo , Regulación hacia Abajo , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Puntos de Control de la Fase M del Ciclo Celular/efectos de los fármacos , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Metaloproteinasa 2 de la Matriz/genética , Metaloproteinasa 9 de la Matriz/genética , Metástasis de la Neoplasia , Inhibidor Tisular de Metaloproteinasa-1/genética , Inhibidor Tisular de Metaloproteinasa-1/metabolismo , Regulación hacia ArribaRESUMEN
Conventional non-surgical therapeutic regimens against osteosarcoma are subject to chemoresistance and tumor relapse, and immunotherapy may be promising for this tumor. However, it's hard to find satisfactory epitopes for immunotherapy against osteosarcoma. Cancer/testis antigens (CTAs), such as MAGE-A family and NY-ESO-1, the potential antigens that almost exclusively express in tumor cells and immune-privileged sites, have been found expressed in osteosarcoma also. Nevertheless, the expression of CTAs is downregulated in many tumors, constraining the application of immunotherapy. In this article, we demonstrate that the expression of MAGE-A family and NY-ESO-1 in osteosarcoma cells can be upregulated following treatment with demethylating agent 5-aza-2'-deoxycytidine and consequently induces a CTA specific CD8+ T-cell response against osteosarcoma in vitro and in vivo. The in vivo imaging was realized by using luciferase-transfected HOS cells and DiR labeled T-cells in severely combined immunodeficiency mouse models. Cytotoxic T cells specifically recognizing MAGE-A family and NY-ESO-1 clustered at the tumor site in mice pre-treated with DAC and resulted in tumor growth suppression, while it was not observed in mice without DAC pre-treatment. This study is important for more targeted therapeutic approaches and suggests that adoptive immunotherapy, combined with demethylating treatment, has the potential for non-surgical therapeutic strategy against osteosarcoma.
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Antígenos de Neoplasias/metabolismo , Azacitidina/análogos & derivados , Neoplasias Óseas/inmunología , Linfocitos T CD8-positivos/inmunología , Proteínas de la Membrana/metabolismo , Proteínas de Neoplasias/metabolismo , Osteosarcoma/inmunología , Linfocitos T Citotóxicos/inmunología , Animales , Antígenos de Neoplasias/genética , Antígenos de Neoplasias/inmunología , Antimetabolitos Antineoplásicos/farmacología , Apoptosis , Azacitidina/farmacología , Western Blotting , Neoplasias Óseas/tratamiento farmacológico , Neoplasias Óseas/patología , Linfocitos T CD8-positivos/efectos de los fármacos , Linfocitos T CD8-positivos/patología , Proliferación Celular , Decitabina , Femenino , Citometría de Flujo , Humanos , Inmunoterapia , Proteínas de la Membrana/genética , Proteínas de la Membrana/inmunología , Ratones , Ratones SCID , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/inmunología , Osteosarcoma/tratamiento farmacológico , Osteosarcoma/patología , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células Tumorales CultivadasRESUMEN
Ethyl(R)-4-chloro-3-hydroxybutanoate ((R)-CHBE) are obtained by cetyltrimetylammonium bromide (CTAB) permeabilized fresh brewer's yeast whole cells bioconversion of ethyl 4-chloro-3-oxobutanoate (COBE ) in the presence of allyl bromide. The results showed that the activities of alcohol dehydrogenase (ADH) and glucose-6-phosphate dehydrogenase (G6PDH) in CTAB permeabilized brewer's yeast cells increased 525 and 7.9-fold, respectively, compared with that in the nonpermeabilized cells and had high enantioselectivity to convert COBE to (R)-CHBE. As one of co-substrates, glucose-6-phosphate was preprepared using glucose phosphorylation by hexokinase-catalyzed of CTAB permeabilized brewer's yeast cells. In a two phase reaction system with n-butyl acetate as organic solvent and with 2-propanol and glucose-6-phosphate as co-substrates, the highest (R)-CHBE concentration of 447 mM was obtained with 110-130 g/l of the CTAB permeabilized cells at optimized pH, temperature, feeding rate and the shake speed of 125 r/min. The yield and enantiomeric excess (ee) of (R)-CHBE reached 99.5 and 99%, respectively, within 6 h.
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Acetoacetatos/metabolismo , Compuestos Alílicos/metabolismo , Permeabilidad de la Membrana Celular , Compuestos de Cetrimonio , Saccharomyces cerevisiae/metabolismo , 2-Propanol/metabolismo , Alcohol Deshidrogenasa/metabolismo , Cetrimonio , Compuestos de Cetrimonio/metabolismo , Glucosa-6-Fosfato/metabolismo , Glucosafosfato Deshidrogenasa/metabolismo , Hexoquinasa/metabolismo , Microbiología Industrial/métodos , NADP/metabolismo , Saccharomyces cerevisiae/crecimiento & desarrolloRESUMEN
OBJECTIVE: To observe ultrastructural changes of the intervertebral disk in the corresponding area after internal fixation of spinal column. METHODS: Twenty-four Japanese big ear rabbits were divided into internal fixation of spinal column group (n=12) and control group (n=12). The internal fixation model was made as follows: The spinous processes and erector spinal muscle were exposed and the T10-L3 spinous processes and the relevant two-side articular processes under the periosteum were isolated. With the help of L-shaped Kirschner wires, the steel wire was threaded through the articular of T11, T12, LI and L2, and were connected with L-shaped Kirschner wires. After 6 months of operation, the following intervertebral disk tissues were observed with transmission electron microscope: nucleus pulposus, internal anulus fibrosus and external anulus fibrosus of L1 intervertebral disk. The T12 and L2 intervertebral disk surface structure was observed horizontally and longitudinally with scanning electron microscope, respectively. RESULTS: After internal fixation of spinal column, the structural changes of cells in nucleus pulposus and internal annulus fibrosus occurred earlier than that in the external annulus fibrosus. Proteoglycan and special structure were found in nucleus pulposus and matrix of annulus fibrosus. However, the forms of special structure in nucleus pulposus and internal layer of annulus fibrosus were different. In the degeneration matrix of intervertebral disc, the proteoglycan particles and special structure were obviously decreased. CONCLUSION: Abnormal stress environment can result in the degeneration of intervertebral disk. There is a regular distribution of the special structure in nucleus pulposus and matrix of annulus fibrosus, which is related to biology behaviour of proteoglycan particles in the degeneration of intervertebral disk.