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1.
J Org Chem ; 89(9): 6027-6039, 2024 May 03.
Artigo em Inglês | MEDLINE | ID: mdl-38630554

RESUMO

In this study, [1+2+2] cyclization of tryptamine-derived isocyanides with 3-ylideneoxindoles was systematically investigated. A series of structurally complex spiro-oxindole derivatives were obtained. Characteristic dynamic covalent chemistry was observed and confirmed by experiments and density functional theory calculation. Through the regulation of the solvent, temperature, and time, the precise and stereodivergent synthesis of spiro-oxindoles was achieved.

2.
Opt Express ; 30(24): 43143-43156, 2022 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-36523019

RESUMO

In 1981, Caves pointed out that the phase sensitivity of a Mach-Zehnder interferometer with single-mode inputs is bounded by the shot-noise limit. The quantum Fisher information analysis shows that this statement holds true for the scenario where two antisymmetric phase shifts occur in two arms, but it is invalid for the scenario where an unknown phase is embedded in one of two arms. In this paper, we focus on the phase sensitivity directed against the latter scenario. The optimal single-mode input is discussed by analyzing common states, including displaced squeezed states, displaced number states, squeezed number states, Schrödinger cat states and completely mixed states. We find that the best choice is a squeezed vacuum state and show the specific measurement scheme which is capable of saturating the corresponding phase sensitivity limit. In addition, we study the effects of several realistic factors-anti-squeezing noise, photon loss and dark counts-on the phase sensitivity. Our results suggest that sub-shot-noise-limited phase sensitivity is attainable with low noise or loss, which paves the way for practical metrology.

3.
BMC Cancer ; 22(1): 691, 2022 Jun 23.
Artigo em Inglês | MEDLINE | ID: mdl-35739510

RESUMO

BACKGROUND: Renal cell carcinoma (RCC) is a third most common tumor of the urinary system. Nowadays, Immunotherapy is a hot topic in the treatment of solid tumors, especially for those tumors with pre-activated immune state. METHODS: In this study, we downloaded genomic and clinical data of RCC samples from The Cancer Genome Atlas (TCGA) database. Four immune-related genetic signatures were used to predict the prognosis of RCC by Cox regression analysis. Then we established a prognostic risk model consisting of the genes most related to prognosis from four signatures to value prognosis of the RCC samples via Kaplan-Meier (KM) survival analysis. An independent data from International Cancer Genome Consortium (ICGC) database were used to test the predictive stability of the model. Furthermore, we performed landscape analysis to assess the difference of gene mutant in the RCC samples from TCGA. Finally, we explored the correlation between the selected genes and the level of tumor immune infiltration via Tumor Immune Estimation Resource (TIMER) platform. RESULTS: We used four genetic signatures to construct prognostic risk models respectively and found that each of the models could divide the RCC samples into high- and low-risk groups with significantly different prognosis, especially in advanced RCC. A comprehensive prognostic risk model was constructed by 8 candidate genes from four signatures (HLA-B, HLA-A, HLA-DRA, IDO1, TAGAP, CIITA, PRF1 and CD8B) dividing the advanced RCC samples from TCGA database into high-risk and low-risk groups with a significant difference in cancer-specific survival (CSS). The stability of the model was verified by independent data from ICGC database. And the classification efficiency of the model was stable for the samples from different subgroups. Landscape analysis showed that mutation ratios of some genes were different between two risk groups. In addition, the expression levels of the selected genes were significantly correlated with the infiltration degree of immune cells in the advanced RCC. CONCLUSIONS: Sum up, eight immune-related genes were screened in our study to construct prognostic risk model with great predictive value for the prognosis of advanced RCC, and the genes were associated with infiltrating immune cells in tumors which have potential to conduct personalized treatment for advanced RCC.


Assuntos
Carcinoma de Células Renais , Neoplasias Renais , Biomarcadores Tumorais/genética , Carcinoma de Células Renais/patologia , Regulação Neoplásica da Expressão Gênica , Humanos , Estimativa de Kaplan-Meier , Neoplasias Renais/patologia , Prognóstico , Fatores de Risco
4.
Chembiochem ; 22(1): 124-128, 2021 01 05.
Artigo em Inglês | MEDLINE | ID: mdl-32789939

RESUMO

Optically active ß-amino alcohols are very useful chiral intermediates frequently used in the preparation of pharmaceutically active substances. Here, a novel cyclohexylamine oxidase (ArCHAO) was identified from the genome sequence of Arthrobacter sp. TYUT010-15 with the R-stereoselective deamination activity of ß-amino alcohol. ArCHAO was cloned and successfully expressed in E. coli BL21, purified and characterized. Substrate-specific analysis revealed that ArCHAO has high activity (4.15 to 6.34 U mg-1 protein) and excellent enantioselectivity toward the tested ß-amino alcohols. By using purified ArCHAO, a wide range of racemic ß-amino alcohols were resolved, (S)-ß-amino alcohols were obtained in >99 % ee. Deracemization of racemic ß-amino alcohols was conducted by ArCHAO-catalyzed enantioselective deamination and transaminase-catalyzed enantioselective amination to afford (S)-ß-amino alcohols in excellent conversion (78-94 %) and enantiomeric excess (>99 %). Preparative-scale deracemization was carried out with 50 mM (6.859 g L-1 ) racemic 2-amino-2-phenylethanol, (S)-2-amino-2-phenylethanol was obtained in 75 % isolated yield and >99 % ee.


Assuntos
Amino Álcoois/metabolismo , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/metabolismo , Transaminases/metabolismo , Amino Álcoois/química , Arthrobacter/enzimologia , Biocatálise , Estrutura Molecular , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/genética , Estereoisomerismo , Transaminases/genética
5.
Mol Biol Rep ; 48(10): 6897-6909, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34453674

RESUMO

BACKGROUND: Environmental hypoxia affects the survival and development of organisms. It is also an important environmental factor that leads to oxidative damage. Hypoxia is a condition in which tissues are deprived of oxygen; reoxygenation is the phenomenon in which hypoxic tissues are exposed to oxygen. Hypoxia-reoxygenation is vital in pathogenesis, where the production of reactive oxygen species and antioxidant disparity significantly contribute to disease progression, and it is one of the most common physiological stressors in the aquaculture industry. METHODS AND RESULTS: In this study, the full length of complementary DNA (cDNA) of the manganese superoxide dismutase (Mn-SOD) gene of healthy cobia Rachycentron canadum was analysed using rapid amplification of cDNA ends. The real-time quantitative Polymerase Chain Reaction was used to measure the expression levels of Mn-SOD mRNAs in various tissues (heart, muscle, brain, liver, kidney, gill, intestine, and spleen). The 2-ΔΔCT method was used to performed the expression analysis. The experimental data were analysed using SPSS ver. 19.0 ( https://spss.software.informer.com/19.0/ ). P < 0.05 and P < 0.01 were set as significant differences. The values were articulated as mean ± standard deviation. The Mn-SOD gene cDNA sequence was 1209 bp long, including a 684 bp open reading frame, 42 bp 5'UTR and 483 bp 3'UTR, encoding 227 amino acids. Under hypoxia-reoxygen stress, the expression of Mn-SOD in brain tissue was significantly lower than in the control group after 8 h of reoxygenation and higher than the control group after 24 h. Hypoxia and subsequent reoxygenation triggered a disturbance in antioxidant homeostasis, displayed in the modification of GPx expression/activity in the liver: GPx was improved. CONCLUSIONS: These results provide valuable information on the role of Mn-SOD regulation in oxidative stress caused by hypoxia.


Assuntos
Antioxidantes/metabolismo , Regulação Enzimológica da Expressão Gênica , Perciformes/genética , Estresse Fisiológico , Superóxido Dismutase/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Hipóxia Celular , Clonagem Molecular , DNA Complementar/genética , Perfilação da Expressão Gênica , Modelos Moleculares , Estresse Oxidativo/genética , Filogenia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Superóxido Dismutase/química
6.
Fish Physiol Biochem ; 47(6): 1951-1967, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34633578

RESUMO

At present, due to the influence of global warming, seasonal change, diurnal variation, and eutrophication of the water body, hypoxia has become one of the major factors limiting the stable development of cobia (Rachycentron canadum) culture. In this study, the miRNAs involved in hypoxia stress were screened, and the target genes of miRNAs were annotated and analyzed. The results showed that a total of 184 conservative microRNA (miRNA) and 121 newly predicted miRNA were obtained by sequencing the liver of control (C) and hypoxic (dissolved oxygen, DO (2.64 ± 0.25) mg/L; 3 h) (S) groups. The pathways involved in energy metabolism included starch and sucrose metabolism (ko00500), glycosaminoglycan degradation (ko00531), and galactose metabolism (ko00052). The results indicate that the body maintains physiological activities by regulating some important pathways at the transcriptional level under hypoxia stress, such as the conversion of aerobic metabolism and anaerobic metabolism, the reduction of energy consumption, and the promotion of red blood cell proliferation to maintain the homeostasis of the body.


Assuntos
Hipóxia , Fígado/metabolismo , MicroRNAs , Perciformes , Animais , Hipóxia/genética , MicroRNAs/genética , Perciformes/genética
7.
Opt Express ; 28(3): 4320-4332, 2020 Feb 03.
Artigo em Inglês | MEDLINE | ID: mdl-32122086

RESUMO

Super-resolved angular displacement estimation is of crucial significance to the field of quantum information processing. Here we report an estimation protocol based on a Sagnac interferometer fed by a coherent state carrying orbital angular momentum. In a lossless scenario, through the use of parity measurement, our protocol can achieve a 4ℓ-fold super-resolved output with quantum number ℓ; meanwhile, a shot-noise-limited sensitivity saturating the quantum Cramér-Rao bound is reachable. We also consider the effects of several realistic factors, including nonideal state preparation, photon loss, and inefficient measurement. Finally, with mean photon number N¯=2.297 and ℓ = 1 taken, we experimentally demonstrate a super-resolved effect of angular displacement with a factor of 7.88.

8.
Biotechnol Lett ; 42(8): 1501-1511, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32219689

RESUMO

OBJECTIVES: To screening of bacteria with cyclic amino alcohol deamination activity for enantioselective synthesis of chiral cyclic ß-amino alcohols. RESULTS: A new strain named Arthrobacter sp. TYUT010-15 with the (R)-selective deamination activity of cyclic ß-amino alcohol has been isolated from nature via a high throughput solid-phase screening method. The reaction conditions of TYUT010-15 were optimized. Using the resting cell of TYUT010-15 as the catalyst, kinetic resolution of trans-2-aminocyclopentanol, trans-2-aminocyclohexanol and cis-1-amino-2-indanol was carried out to afford (1S, 2S)-trans-2-aminocyclopentanol, (1S, 2S)-trans-2-aminocyclohexanol and (1R, 2S)-cis-1-amino-2-indanol in > 99% ee and 49.6-50% conversion. Four aromatic ß-amino alcohols and two amines were also resolved, (S)-ß-amino alcohols and (R)-amines were obtained in > 99% ee. Preparation experiment was conducted with 200 mM (23.2 g L-1) racemic trans-2-aminocyclohexanol, yielding the desired (1S, 2S)-trans-2-aminocyclohexanol in 40% isolated yield, > 99% ee and 5.8 g L-1 d-1 space time yields. CONCLUSIONS: This study provides a high throughput solid-phase method for screening of bacteria with cyclic amino alcohol deamination activity and a first example for practical preparation of chiral cyclic ß-amino alcohol by Arthrobacter sp. TYUT010-15.


Assuntos
Amino Álcoois , Bactérias/metabolismo , Ensaios de Triagem em Larga Escala/métodos , Aminas/análise , Aminas/química , Aminas/metabolismo , Amino Álcoois/análise , Amino Álcoois/química , Amino Álcoois/metabolismo , Arthrobacter/genética , Arthrobacter/metabolismo , Bactérias/genética , Colorimetria , Desaminação , Cinética , Estereoisomerismo , Especificidade por Substrato
9.
Opt Express ; 27(4): 5512-5522, 2019 Feb 18.
Artigo em Inglês | MEDLINE | ID: mdl-30876153

RESUMO

We demonstrate a tried-and-true binary strategy for angular displacement estimation, of which the measuring system is a modified Mach-Zehnder interferometer fed by a coherent state carrying orbital angular momentum, and two Dove prisms are embedded in two arms. Unlike previous protocols, in this paper, we use fidelity instead of standard deviation to evaluate the detection strategies. Two binary strategy candidates, parity detection and Z detection, are considered and compared. In addition, we study the effects of several realistic scenarios on the estimation protocol, including transmission loss, detection efficiency, dark counts, and those which are a combination thereof. Finally, we exhibit a proof-of-principle experiment, the results suggest a resolution enhancement effect with a factor of 3.72.

10.
J Am Soc Nephrol ; 29(3): 919-935, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-29295871

RESUMO

Albuminuria is a key instigator of tubulointerstitial inflammation associated with CKD, but the mechanism through which filtered albumin propagates renal injury remains unclear. In this study, we explored the role in this process of exosome mRNA released from tubular epithelial cells (TECs). Compared with control mice, acute and chronic kidney injury models had more exosomes containing inflammatory cytokine mRNA, particularly the chemokine CCL2, in kidneys and urine. In vitro stimulation of TECs with BSA recapitulated this finding. Notably, the internalization of purified TEC exosomes by cultured macrophages increased if TECs were exposed to BSA. Macrophage internalization of exosomes from BSA-treated TECs led to an enhanced inflammatory response and macrophage migration, but CCL2 silencing in TECs prevented these effects. Using a GFP-CCL2 fusion mRNA construct, we observed direct transfer of CCL2 mRNA from TEC exosomes to macrophages. Mice subjected to tail vein injection of purified BSA-treated TEC exosomes developed tubular injury with renal inflammatory cell infiltration. However, injection of exosomes from BSA-treated CCL2-deficient TECs induced less severe kidney inflammation. Finally, in patients with IgA nephropathy, the increase of proteinuria correlated with augmented urinary excretion of exosomes with exaggerated expression of CCL2 mRNA. Moreover, the level of CCL2 mRNA in urinary exosomes correlated closely with levels of renal interstitial macrophage infiltration in these patients. Our studies demonstrate that the increasing release of exosomes that transfer CCL2 mRNA from TECs to macrophages constitutes a critical mechanism of albumin-induced tubulointerstitial inflammation.


Assuntos
Injúria Renal Aguda/metabolismo , Quimiocina CCL2/genética , Quimiocina CCL2/metabolismo , Células Epiteliais/metabolismo , Exossomos/metabolismo , Glomerulonefrite por IGA/urina , Túbulos Renais/metabolismo , Macrófagos/metabolismo , RNA Mensageiro/metabolismo , Insuficiência Renal Crônica/metabolismo , Injúria Renal Aguda/genética , Injúria Renal Aguda/urina , Adulto , Animais , Movimento Celular/efeitos dos fármacos , Células Cultivadas , Modelos Animais de Doenças , Exossomos/genética , Feminino , Inativação Gênica , Glomerulonefrite por IGA/complicações , Glomerulonefrite por IGA/patologia , Humanos , Túbulos Renais/citologia , Túbulos Renais/patologia , Macrófagos/fisiologia , Masculino , Camundongos , Pessoa de Meia-Idade , Nefrite/metabolismo , Nefrite/patologia , Proteinúria/etiologia , Proteinúria/patologia , Proteinúria/urina , Ratos , Insuficiência Renal Crônica/genética , Insuficiência Renal Crônica/urina , Soroalbumina Bovina/farmacologia , Adulto Jovem
11.
World J Microbiol Biotechnol ; 35(11): 179, 2019 Nov 09.
Artigo em Inglês | MEDLINE | ID: mdl-31707486

RESUMO

High concentration restaurant oily wastewater from restaurants and food processing industries discharged into water environment usually results in environment pollution and inhibits the activity of microorganisms in biological wastewater treatment systems. In this study, 75 strains from oily sludge were isolated with oil degradation activity for edible oil-contained wastewater. Eight isolates were able to grow well in liquid cultures with edible oil as the sole carbon source and discovered with high efficient oil-degrading ability. Seven out of eight isolates were identified as Acinetobacter and one isolate as Kluyvera cryocrescens, based on their 16S rRNA gene sequences. Three highly efficient oil degrading bacteria (Acinetobacter dijkshoorniae LYC46-2, Kluyvera cryocrescens LYC50-1a and Acinetobacter pittii LYC73-4b) were selected and their degradation characteristic were examined, the results showed that the three isolates were effective under pH range from 7.0 to 10.0, and temperature from 25 to 35 °C. For degradation of 2-4% (v/v) of vegetable oil, > 85% degradation percentage were obtained within 30 h. Degradation of the higher concentration oil (6-8%, v/v) result in 50-70% degradation percentage within 72 h, and the degradation percentage for the isolated strains were decreased about 50% for the degradation of 10% oil (< 45%) compared to 2% oil. Different type of oils were also tested, > 90% of degradation percentage were obtained by the three isolates, implied that these strains are capable of removing various oils efficiently. These results suggested that Acinetobacter dijkshoorniae LYC46-2, Kluyvera cryocrescens LYC50-1a and Acinetobacter pittii LYC73-4b are potential species could be efficiently used for high concentration restaurant oily wastewater treatment and might be applicable to a wastewater treatment system for the removal of oil.


Assuntos
Bactérias/isolamento & purificação , Restaurantes , Esgotos/microbiologia , Águas Residuárias/microbiologia , Acinetobacter/genética , Acinetobacter/isolamento & purificação , Bactérias/genética , Bactérias/crescimento & desenvolvimento , Biodegradação Ambiental , Concentração de Íons de Hidrogênio , Kluyvera/genética , Kluyvera/isolamento & purificação , Filogenia , Óleos de Plantas , RNA Ribossômico 16S/genética , Temperatura
12.
Opt Express ; 26(13): 16524-16534, 2018 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-30119481

RESUMO

We report on an orbital-angular-momentum-enhanced scheme for angular displacement estimation based on two-mode squeezed vacuum and parity detection. The sub-Heisenberg-limited sensitivity for angular displacement estimation is obtained in an ideal situation. Several realistic factors are also considered, including photon loss, dark counts, response-time delay, and thermal photon noise. Our results indicate that the effects of realistic factors on the sensitivity can be offset by raising orbital angular momentum quantum number ℓ. This implies that the robustness and the practicability of the system can be improved via raising ℓ without changing mean photon number N.

13.
Opt Express ; 26(25): 33080-33090, 2018 Dec 10.
Artigo em Inglês | MEDLINE | ID: mdl-30645465

RESUMO

In this paper, we propose a protocol for the estimation of angular displacement based upon orbital angular momentum and an SU(1,1)-SU(2) hybrid interferometer. This interferometer consists of an optical parametric amplifier, a beam splitter, and reflection mirrors; the balanced homodyne detection is used as the detection strategy. The results indicate that super-resolution and super-sensitivity can be achieved with an ideal scenario. Additionally, we study the effect of photon loss on resolution and sensitivity, and the robustness of our protocol is also discussed. Finally, the advantage of our protocol compared with an SU(1,1) protocol is demonstrated, and the merits of orbital angular momentum-enhanced protocol are summarized.

14.
Biotechnol Lett ; 40(2): 349-358, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29124518

RESUMO

OBJECTIVES: To investigate the efficiency of a new cascade biocatalysis system for the conversion of R, S-ß-amino alcohols to enantiopure vicinal diol and ß-amino alcohol. RESULTS: An efficient cascade biocatalysis was achieved by combination of a transaminase, a carbonyl reductase and a cofactor regeneration system. An ee value of > 99% for 2-amino-2-phenylethanol and 1-phenyl-1, 2-ethanediol were simultaneously obtained with 50% conversion from R, S-2-amino-2-phenylethanol. The generality of the cascade biocatalysis was further demonstrated with the whole-cell approaches to convert 10-60 mM R, S-ß-amino alcohol to (R)- and (S)-diol and (R)- and (S)-ß-amino alcohol in 90-99% ee with 50-52% conversion. Preparative biotransformation was demonstrated at a 50 ml scale with mixed recombinant cells to give both (R)- and (S)-2-amino-2-phenylethanol and (R)- and (S)-1-phenyl-1, 2-ethanediol in > 99% ee and 40-42% isolated yield from racemic 2-amino-2-phenylethanol. CONCLUSIONS: This cascade biocatalysis system provides a new practical method for the simultaneous synthesis of optically pure vicinal diol and an ß-amino alcohol.


Assuntos
Oxirredutases do Álcool/metabolismo , Amino Álcoois/química , Amino Álcoois/metabolismo , Biotecnologia/métodos , Amino Álcoois/análise , Proteínas de Bactérias/metabolismo , Biocatálise , Sistema Livre de Células , Escherichia coli/enzimologia , Estereoisomerismo , Transaminases/metabolismo
15.
Am J Physiol Renal Physiol ; 312(3): F436-F444, 2017 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-27582099

RESUMO

Previous studies have shown that increased parathyroid hormone (PTH) attributable to secondary hyperparathyroidism in chronic kidney disease accelerates the arteriosclerotic fibrosis and calcification. Although the underlying mechanisms remain largely unknown, endothelial cells (ECs) have recently been demonstrated to participate in calcification in part by providing chondrogenic cells via the endothelial-to-mesenchymal transition (EndMT). Therefore, this study aimed to investigate whether elevated PTH could induce endothelial-to-chondrogenic transition in aortic ECs and to determine the possible underlying signaling pathway. We found that treatment of ECs with PTH significantly upregulated the expression of EndMT-related markers. Accordingly, ECs treated with PTH exhibited chondrogenic potential. In vivo, lineage-tracing model-subjected mice with endothelial-specific green fluorescent protein fluorescence to chronic PTH infusion showed a marked increase in the aortic expression of chondrocyte markers, and confocal microscopy revealed the endothelial origin of cells expressing chondrocyte markers in the aorta after PTH infusion. Furthermore, this in vitro study showed that PTH enhanced the nuclear localization of ß-catenin in ECs, whereas ß-catenin siRNA or DKK1, an inhibitor of ß-catenin nuclear translocation, attenuated the upregulation of EndMT-associated and chondrogenic markers induced by PTH. In summary, our study demonstrated that elevated PTH could induce the transition of ECs to chondrogenic cells via EndMT, possibly mediated by the nuclear translocation of ß-catenin.


Assuntos
Aorta/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Condrogênese/efeitos dos fármacos , Células Endoteliais/efeitos dos fármacos , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Hormônio Paratireóideo/farmacologia , Transporte Ativo do Núcleo Celular , Animais , Aorta/metabolismo , Aorta/patologia , Linhagem da Célula , Células Cultivadas , Relação Dose-Resposta a Droga , Células Endoteliais/metabolismo , Células Endoteliais/patologia , Humanos , Camundongos Transgênicos , Fenótipo , Interferência de RNA , Transdução de Sinais/efeitos dos fármacos , Transfecção , beta Catenina/genética , beta Catenina/metabolismo
16.
Am J Pathol ; 186(11): 2846-2856, 2016 11.
Artigo em Inglês | MEDLINE | ID: mdl-27640148

RESUMO

Inappropriate activation of the renin angiotensin system (RAS) is a key contributor to the pathogenesis of essential hypertension. During RAS activation, infiltration of immune cells into the kidney exacerbates hypertension and renal injury. However, the mechanisms underpinning the accumulation of mononuclear cells in the kidney after RAS stimulation remain unclear. C-C motif chemokine 5 (CCL5) drives recruitment of macrophages and T lymphocytes into injured tissues, and we have found that RAS activation induces CCL5 expression in the kidney during the pathogenesis of hypertension and renal fibrosis. We therefore evaluated the contribution of CCL5 to renal damage and fibrosis in hypertensive and normotensive models of RAS stimulation. Surprisingly, during angiotensin II-induced hypertension, CCL5-deficient (knockout, KO) mice exhibited markedly augmented kidney damage, macrophage infiltration, and expression of proinflammatory macrophage cytokines compared with wild-type controls. When subjected to the normotensive unilateral ureteral obstruction model of endogenous RAS activation, CCL5 KO mice similarly developed more severe renal fibrosis and greater accumulation of macrophages in the kidney, congruent with enhanced renal expression of the macrophage chemokine CCL2. In turn, pharmacologic inhibition of CCL2 abrogated the differences between CCL5 KO and wild-type mice in kidney fibrosis and macrophage infiltration after unilateral ureteral obstruction. These data indicate that CCL5 paradoxically limits macrophage accumulation in the injured kidney during RAS activation by constraining the proinflammatory actions of CCL2.


Assuntos
Angiotensina II/imunologia , Quimiocina CCL5/metabolismo , Hipertensão/imunologia , Nefropatias/imunologia , Rim/patologia , Animais , Pressão Sanguínea , Quimiocina CCL2/genética , Quimiocina CCL2/metabolismo , Quimiocina CCL5/genética , Hipertensão Essencial , Feminino , Fibrose , Hipertensão/etiologia , Rim/imunologia , Rim/cirurgia , Nefropatias/etiologia , Macrófagos/imunologia , Macrófagos/metabolismo , Masculino , Camundongos , Camundongos Knockout , Nefrectomia , Sistema Renina-Angiotensina/imunologia , Linfócitos T/imunologia , Obstrução Ureteral
17.
Anal Biochem ; 518: 94-101, 2017 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-27899283

RESUMO

Chiral vicinal amino alcohols are important chiral building blocks and intermediates in the pharmaceutical industry. The transaminase (TAm) catalyzed kinetic resolution of racemic amino alcohols provides a straightforward approach to access these important compounds. This study describes the development of a novel microtiter plate assay to screen vicinal amino alcohol-specific TAms using a tetrazolium red-based colorimetric assay to monitor the rate of α-hydroxy ketone formation at 510 nm. This approach is the first to determine the Michaelis-Menten parameters for a recombinant TAm (PpbauA) from Pseudomonas putida NBRC14164. The corresponding Vmax and KM values for both enantiomers of 2-amino-1-propanol and 2-amino-1-butanol were obtained, and the calculated kinetic E-factors of PpbauA toward 2-amino-1-propanol and 2-amino-1-butanol are 3 (S) and 6 (R), respectively. The method is sensitive and exhibits low level background coloration. Moreover, this method can be used to detect transaminase activity and enantioselectivity toward amino alcohols in a high-throughput format. Additionally, this simple method is compatible with the most widely used (R)- and (S)-selective transaminases and may be a broadly applicable tool for screening transaminases from a transaminase mutant library.


Assuntos
Amino Álcoois/química , Proteínas de Bactérias/química , Propanolaminas/química , Pseudomonas putida/enzimologia , Transaminases/química , Amino Álcoois/metabolismo , Proteínas de Bactérias/metabolismo , Propanolaminas/metabolismo , Especificidade por Substrato/fisiologia , Transaminases/metabolismo
18.
Tumour Biol ; 37(3): 3797-805, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26472724

RESUMO

Delta-like ligand-4 (DLL4)-Notch signaling is known to play a pivotal role in the regulation of tumor angiogenesis. We had previously found that DLL4 was overexpressed, while Notch1 receptor, which binds to DLL4 during angiogenesis, was absent in the majority of human primary glioblastomas. Thus, DLL4-Notch signaling pathway in the regulation of tumor angiogenesis in primary glioblastoma remains unknown. Tumor tissues from 70 patients with primary glioblastoma were analyzed by immunohistochemistry for expression of components of DLL4-Notch signaling, vascular endothelial growth factor (VEGF), and microvessel density (MVD). Immunohistochemistry results showed that the positive staining of DLL4 and Notch4 was primarily distributed in tumor vascular endothelial cells but rarely detected in tumor cells. However, VEGF, hairy/enhancer of split-1 (HES1; a target gene of Notch signaling), and Notch1-3 expression was seen in both tumor vascular endothelial cells and tumor cells. Univariate analysis showed that the expression levels of VEGF and DLL4, HES1, and Notch4 in tumor endothelial cells were significantly associated with MVD in primary glioblastoma (P < 0.001). Binary logistic regression analysis showed that high expression levels of DLL4, HES1, and Notch4 in tumor endothelial cells were associated with a decrease of MVD in primary glioblastoma, while MVD increased with elevated VEGF expression in contrast. In addition, DLL4, Notch4, and HES1 expression were positively correlated in tumor vascular endothelial cells (P < 0.05). We conclude that the vascular DLL4-Notch4 signaling and VEGF signaling complementing each other plays an important role in the progression of tumor angiogenesis in primary glioblastoma. Graphical abstract A, positive staining of DLL4 in human kidney; B, positive staining of VEGF in human breast cancer; C, positive staining of CD34 in human lung cancer; D, positive staining of HES1 in human breast cancer; E-H, positive staining of Notch1-4: E-F in human lung cancer; G-H in human kidney.


Assuntos
Glioblastoma/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/biossíntese , Proteínas de Membrana/biossíntese , Neovascularização Patológica/metabolismo , Proteínas Proto-Oncogênicas/biossíntese , Receptores Notch/biossíntese , Transdução de Sinais , Adolescente , Adulto , Idoso , Feminino , Glioblastoma/irrigação sanguínea , Glioblastoma/patologia , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Receptor Notch1/biossíntese , Receptor Notch2/biossíntese , Receptor Notch3/biossíntese , Receptor Notch4 , Fatores de Transcrição HES-1/biossíntese , Fator A de Crescimento do Endotélio Vascular/biossíntese , Adulto Jovem
19.
Bioprocess Biosyst Eng ; 39(4): 603-11, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26801669

RESUMO

Two uncharacterized nicotinamide adenine dinucleotide (NADH) oxidases (named as LpNox1, LpNox2) from Lactobacillus pentosus ATCC 8041 were cloned and overexpressed in Escherichia coli BL21 (DE3). The sequence analysis revealed that the two enzymes are water-forming Noxs with 64 % and 52 % identity to LbNox from Lactobacillus brevis DSM 20054. The optimal pH and temperature of the purified LpNox1 and LpNox2 were 7.0 and 8.0 and 35 and 40 °C, respectively, with K M of 99.0 µM (LpNox1) and 27.6 µM (LpNox2), and yielding catalytic efficiency k cat/K M of 1.0 and 0.2 µM(-1) s(-1), respectively. Heat inactivation studies revealed that the two enzymes are relatively instable. The application of LpNox1 for the regeneration of NAD(+) was demonstrated by coupling with a glycerol dehydrogenase-catalyzed oxidation of glycerol to 1,3-dihydroxyacetone. The characteristics of the LpNox1 could prove to be of interest in industrial application such as NAD(+) regeneration in dehydrogenase-catalyzed oxidations.


Assuntos
Proteínas de Bactérias , Lactobacillus pentosus , NADPH Oxidases , NAD/metabolismo , Proteínas de Bactérias/biossíntese , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Catálise , Clonagem Molecular , Concentração de Íons de Hidrogênio , Lactobacillus pentosus/enzimologia , Lactobacillus pentosus/genética , NAD/genética , NADPH Oxidases/biossíntese , NADPH Oxidases/química , NADPH Oxidases/genética , Oxirredução
20.
Zhonghua Nan Ke Xue ; 22(10): 914-922, 2016 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-29278474

RESUMO

OBJECTIVE: To evaluate the safety and effect of transurethral holmium laser enucleation of the prostate (HoLEP) in comparison with bipolar transurethral plasmakinetic prostatectomy (TUPKP) in the treatment of benign prostatic hyperplasia (BPH). METHODS: We searched the databases of PubMed, SCI, Ovid, The Cochrane Library, CNKI, CBM, VIP, and Wangfang Data for controlled clinical trials about HoLEP versus TUPKP in the treatment of BPH published up to April 2016. The studies were screened according to the inclusion and exclusion criteria, the data extracted, and their quality evaluated by 2 reviewers independently, followed by a meta-analysis using the RevMan 5.3 software. RESULTS: A total of 7 studies were included, involving 2031 cases. In comparison with TUPKP, HoLEP showed significantly longer operation time (WMD = 24.61, 95% CI 11.88, 37.34, P lt; 0.001), shorter hospital stay (WMD =-1.91, 95% CI -3.74, -0.07, P = 0.04), shorter bladder irrigation time (WMD = -21.50, 95% CI -34.95, -8.06, P = 0.002), shorter catheter-indwelling time (WMD = -27.60, 95% CI -48.17, -7.03, P = 0.009), less hemoglobin loss (WMD = - 0.42, 95% CI -0.78, -0.07, P = 0.02); lower postvoid residual urine (PVR) at 3 months (WMD = -3.35, 95% CI -4.46, -2.23, P<0.001) and 6 months after surgery (WMD =-1.11, 95% CI -2.18, -0.05, P = 0.04); higher maximum urinary flow rate (Qmax) (WMD = 0.42, 95% CI 0.04, 0.80, P = 0.03) and fewer urinary tract irritation symptoms (OR =0.58, 95% CI 0.41, 0.81, P = 0.002) at 12 months after surgery. No statistically significant differences were found between the two groups in the volume of resected tissue, serum sodium reduction, urethral stricture, erectile dysfunction, retrograde ejaculation, or transient urinary incontinence (P>0.05), or in the improvement of the quality of life (QoL) at 1, 3 and 12 months, International Prostate Symptom Score (IPSS) at 1, 3, 6 and 12 months, Qmax at 1, 3 and 6 months, or International Index of Erectile Function-5 (IIEF-5) at 6 months after surgery (P>0.05). CONCLUSIONS: HoLEP is preferred to TUPKP in clinical application for its advantages of higher Qmax at 12 months after surgery, lower PVR at 3 and 6 months, higher peri-operative safety, faster recovery, and fewer urinary tract irritation symptoms. However, for the quantity and quality limitations of the included publications, our findings are to be further supported by large-sample, multi-center, and high-quality prospective controlled clinical studies.


Assuntos
Terapia a Laser , Prostatectomia/métodos , Hiperplasia Prostática/cirurgia , Ressecção Transuretral da Próstata , Disfunção Erétil , Hólmio , Humanos , Lasers de Estado Sólido , Tempo de Internação , Masculino , Qualidade de Vida , Estreitamento Uretral , Bexiga Urinária , Incontinência Urinária , Retenção Urinária
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