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1.
Genomics ; 115(4): 110662, 2023 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-37270068

RESUMEN

cfDNA fragmentomic features have been used in cancer detection models; however, the generalizability of the models needs to be tested. We proposed a type of cfDNA fragmentomic feature named chromosomal arm-level fragment size distribution (ARM-FSD), evaluated and compared its performance and generalizability for lung cancer and pan-cancer detection with existing cfDNA fragmentomic features (as reference) by using cohorts from different institutions. The ARM-FSD lung cancer model outperformed the reference model by ∼10% when being tested by two external cohorts (AUC: 0.97 vs. 0.86; 0.87 vs. 0.76). For pan-cancer detection, the performance of the ARM-FSD based model is consistently higher than the reference (AUC: 0.88 vs. 0.75, 0.98 vs. 0.63) in a pan-cancer and a lung cancer external validation cohort, indicating that ARM-FSD model produces stable performance across multiple cohorts. Our study reveals ARM-FSD based models have a higher generalizability, and highlights the necessity of cross-study validation for predictive model development.


Asunto(s)
Ácidos Nucleicos Libres de Células , Trastornos de los Cromosomas , Neoplasias Pulmonares , Humanos , Ácidos Nucleicos Libres de Células/genética , Detección Precoz del Cáncer , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/genética , Trastornos de los Cromosomas/diagnóstico , Biomarcadores de Tumor/genética
2.
Clin Chem ; 69(1): 88-99, 2023 01 04.
Artículo en Inglés | MEDLINE | ID: mdl-36308331

RESUMEN

BACKGROUND: A "Watch and Wait" (W&W) approach has become an alternative to surgery for locally advanced rectal cancer (LARC) after neoadjuvant chemoradiotherapy (nCRT). Precise prediction of pathological complete response (pCR) will improve patient selection for W&W. We investigated the utility of cell-free DNA (cfDNA) fragmentomics in predicting pCR. METHODS: We recruited 119 LARC patients and evaluated nCRT response by pCR status and pathological or MRI tumor regression grade (mrTRG). Plasma samples before, during, and after nCRT were applied to deep targeted-panel sequencing, with 103 patients having complete samples. cfDNA fragment and 5'-end motif profiles were used to construct elastic-net logistic regression models to predict non-pCR. Predictive performance was measured by area under the receiver operator characteristic curve (AUC), sensitivity, and specificity. RESULTS: In the training cohort, the model based on 5'-end motif profile plus mrTRG achieved the highest cross-validation AUC (0.92, 95% CI, 0.91-0.93). The AUC in a testing cohort was 0.96 (95% CI, 0.90-1.00). The models based on 5'-end motif profile alone or in combination with mrTRG both maintained good predictive ability for patients without detectable circulating tumor DNA (AUC 0.94, 95% CI, 0.93-0.95; AUC 0.95, 95% CI, 0.94-0.96). In an external validation cohort, the model trained with a local 5'-end motif profile obtained an AUC of 0.878 (95% CI, 0.801-0.956) in discriminating colorectal cancer from healthy subjects. CONCLUSIONS: The combination of a 5'-end motif profile with mrTRG has the potential to predict the response to nCRT, and therefore may improve the patient selection for a W&W approach.


Asunto(s)
Ácidos Nucleicos Libres de Células , Neoplasias del Recto , Humanos , Neoplasias del Recto/terapia , Neoplasias del Recto/tratamiento farmacológico , Terapia Neoadyuvante , Ácidos Nucleicos Libres de Células/genética , Resultado del Tratamiento , Quimioradioterapia , ADN , Estudios Retrospectivos
3.
Blood Press ; 32(1): 2195009, 2023 12.
Artículo en Inglés | MEDLINE | ID: mdl-37020399

RESUMEN

Purpose: Reduced slow wave sleep (SWS) has been linked to hypertension in some studies. The aim of the study is to investigate the association between SWS and office blood pressure (BP) in non-hypertensive obstructive sleep apnea (OSA).Methods: This is a retrospective study of 3350 patients who underwent polysomnography (PSG) in our hospital. Based on quartiles of percent SWS, participants were classified into four groups. BP was measured manually on the randomly chosen arm in a seated position with sphygmomanometer after PSG in the morning, and the average of the second and third measurements was used for this analysis. Elevated office BP was defined as a systolic BP≥140 mmHg or diastolic BP≥90 mmHg.Results: There were 1365 patients with OSA and 597 primary snorers included in our study. In OSA group, OSA patients with SWS <13.5% had a significant elevated risk with elevated office BP (OR,1.49[95%CI 1.05-2.10], P=0.025), compared to the highest quartile (percent SWS >39.2%). However, no significant relationship between decreased SWS and elevated office BP was found in primary snorers group.Conclusion: In non-hypertensive OSA patients, decreased SWS is associated with elevated office BP.


This is the first study to investigate the association between decreased SWS and incident elevated office BP in non-hypertensive OSA patients.Our results found that in non-hypertensive OSA patients, decreased SWS is associated with elevated office BP.The relationship between decreased SWS and elevated office BP in OSA patients was evident especially in men and in those <60 years old.


Asunto(s)
Hipertensión , Apnea Obstructiva del Sueño , Sueño de Onda Lenta , Humanos , Presión Sanguínea/fisiología , Estudios Transversales , Estudios Retrospectivos , Apnea Obstructiva del Sueño/complicaciones , Sueño
4.
PLoS Med ; 18(8): e1003741, 2021 08.
Artículo en Inglés | MEDLINE | ID: mdl-34464382

RESUMEN

BACKGROUND: For locally advanced rectal cancer (LARC) patients who receive neoadjuvant chemoradiotherapy (nCRT), there are no reliable indicators to accurately predict pathological complete response (pCR) before surgery. For patients with clinical complete response (cCR), a "Watch and Wait" (W&W) approach can be adopted to improve quality of life. However, W&W approach may increase the recurrence risk in patients who are judged to be cCR but have minimal residual disease (MRD). Magnetic resonance imaging (MRI) is a major tool to evaluate response to nCRT; however, its ability to predict pCR needs to be improved. In this prospective cohort study, we explored the value of circulating tumor DNA (ctDNA) in combination with MRI in the prediction of pCR before surgery and investigated the utility of ctDNA in risk stratification and prognostic prediction for patients undergoing nCRT and total mesorectal excision (TME). METHODS AND FINDINGS: We recruited 119 Chinese LARC patients (cT3-4/N0-2/M0; median age of 57; 85 males) who were treated with nCRT plus TME at Fudan University Shanghai Cancer Center (China) from February 7, 2016 to October 31, 2017. Plasma samples at baseline, during nCRT, and after surgery were collected. A total of 531 plasma samples were collected and subjected to deep targeted panel sequencing of 422 cancer-related genes. The association among ctDNA status, treatment response, and prognosis was analyzed. The performance of ctDNA alone, MRI alone, and combining ctDNA with MRI was evaluated for their ability to predict pCR/non-pCR. Ranging from complete tumor regression (pathological tumor regression grade 0; pTRG0) to poor regression (pTRG3), the ctDNA clearance rate during nCRT showed a significant decreasing trend (95.7%, 77.8%, 71.1%, and 66.7% in pTRG 0, 1, 2, and 3 groups, respectively, P = 0.008), while the detection rate of acquired mutations in ctDNA showed an increasing trend (3.8%, 8.3%, 19.2%, and 23.1% in pTRG 0, 1, 2, and 3 groups, respectively, P = 0.02). Univariable logistic regression showed that ctDNA clearance was associated with a low probability of non-pCR (odds ratio = 0.11, 95% confidence interval [95% CI] = 0.01 to 0.6, P = 0.04). A risk score predictive model, which incorporated both ctDNA (i.e., features of baseline ctDNA, ctDNA clearance, and acquired mutation status) and MRI tumor regression grade (mrTRG), was developed and demonstrated improved performance in predicting pCR/non-pCR (area under the curve [AUC] = 0.886, 95% CI = 0.810 to 0.962) compared with models derived from only ctDNA (AUC = 0.818, 95% CI = 0.725 to 0.912) or only mrTRG (AUC = 0.729, 95% CI = 0.641 to 0.816). The detection of potential colorectal cancer (CRC) driver genes in ctDNA after nCRT indicated a significantly worse recurrence-free survival (RFS) (hazard ratio [HR] = 9.29, 95% CI = 3.74 to 23.10, P < 0.001). Patients with detectable driver mutations and positive high-risk feature (HR_feature) after surgery had the highest recurrence risk (HR = 90.29, 95% CI = 17.01 to 479.26, P < 0.001). Limitations include relatively small sample size, lack of independent external validation, no serial ctDNA testing after surgery, and a relatively short follow-up period. CONCLUSIONS: The model combining ctDNA and MRI improved the predictive performance compared with the models derived from individual information, and combining ctDNA with HR_feature can stratify patients with a high risk of recurrence. Therefore, ctDNA can supplement MRI to better predict nCRT response, and it could potentially help patient selection for nonoperative management and guide the treatment strategy for those with different recurrence risks.


Asunto(s)
ADN Tumoral Circulante/uso terapéutico , Terapia Neoadyuvante/estadística & datos numéricos , Neoplasias del Recto/prevención & control , Femenino , Humanos , Masculino , Persona de Mediana Edad , Pronóstico , Estudios Prospectivos , Neoplasias del Recto/diagnóstico , Resultado del Tratamiento
5.
J Recept Signal Transduct Res ; 41(1): 67-73, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-32635793

RESUMEN

Preeclampsia (PE) is a specific disorder of pregnancy with significant morbidity and mortality to the mother and the fetus. It has been reported that abnormal regulation of trophoblast cells contributes to the development of PE. LIM and SH3 Protein 2 (LASP2) belongs to nebulin protein family of actin-binding protein that has broad biological functions. In this study, we evaluated the role of LASP2 in the regulation of trophoblast cells. Our results demonstrated that LASP2 was markedly up-regulated in the placentas of patients with PE. Overexpression of LASP2 significantly suppressed the cell proliferation, migration, and invasion of trophoblast cells. LASP2 overexpression reduced the expression levels of ß-catenin, cyclin D1, and c-Myc in trophoblast cells. Furthermore, activation of Wnt/ß-catenin pathway booked the effects of LASP2 on trophoblast cells. In conclusion, these findings indicated that LASP2 might contribute to the pathogenesis of PE via regulating cell proliferation, migration, and invasion of trophoblast cells. Thus, LASP2 might be a prospective therapeutic target for the prevention and treatment of PE.


Asunto(s)
Proteínas Portadoras/genética , Proliferación Celular/genética , Proteínas del Citoesqueleto/genética , Proteínas con Dominio LIM/genética , Preeclampsia/genética , beta Catenina/genética , Adulto , Línea Celular Tumoral , Movimiento Celular/genética , Ciclina D1/genética , Femenino , Humanos , Placenta/metabolismo , Placenta/patología , Preeclampsia/patología , Embarazo , Proteínas Proto-Oncogénicas c-myc/genética , Trofoblastos/metabolismo , Trofoblastos/patología , Vía de Señalización Wnt/genética
6.
Indoor Air ; 31(6): 2142-2157, 2021 11.
Artículo en Inglés | MEDLINE | ID: mdl-34337798

RESUMEN

Humans emit carbon dioxide (CO2 ) as a product of their metabolism. Its concentration in buildings is used as a marker of ventilation rate (VR) and degree of mixing of supply air, and indoor air quality (IAQ). The CO2 emission rate (CER) may be used to estimate the ventilation rate. Many studies have measured CERs from subjects who were awake but little data are available from sleeping subjects and the present publication was intended to reduce this gap in knowledge. Seven females (29 ± 5 years old; BMI: 22.2 ± 0.8 kg/m2 ) and four males (27 ± 1 years old; BMI: 20.5 ± 1.5 kg/m2 ) slept for four consecutive nights in a specially constructed capsule at two temperatures (24 and 28°C) and two VRs that maintained CO2  levels at ca. 800 ppm and 1700 ppm simulating sleeping conditions reported in the literature. The order of exposure was balanced, and the first night was for adaptation. Their physiological responses, including heart rate, pNN50, core body temperature, and skin temperature, were measured as well as sleep quality, and subjective responses were collected each evening and morning. Measured steady-state CO2 concentrations during sleep were used to estimate CERs with a mass-balance equation. The average CER was 11.0 ± 1.4 L/h per person and was 8% higher for males than for females (P < 0.05). Increasing the temperature or decreasing IAQ by decreasing VR had no effects on measured CERs and caused no observable differences in physiological responses. We also calculated CERs for sleeping subjects using the published data on sleep energy expenditure (SEE) and Respiratory Quotient (RQ), and our measured CERs confirmed both these calculations and the CERs predicted using the equations provided by ASHRAE Standard 62.1, ASHRAE Handbook, and ASTM D6245-18. The present results provide a valuable and helpful reference for the design and control of bedroom ventilation but require confirmation and extension to other age groups and populations.


Asunto(s)
Contaminación del Aire Interior , Dióxido de Carbono , Adulto , Contaminación del Aire Interior/análisis , Dióxido de Carbono/análisis , Femenino , Humanos , Masculino , Sueño , Temperatura , Ventilación , Adulto Joven
7.
Cancer Sci ; 111(6): 2163-2173, 2020 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-32187423

RESUMEN

Papillary thyroid carcinoma (PTC) is the most common malignancy of the thyroid gland, with a relatively high cure rate. Distant metastasis (DM) of PTC is uncommon, but when it occurs, it significantly decreases the survival of PTC patients. The molecular mechanisms of DM in PTC have not been systematically studied. We performed whole exome sequencing and GeneseeqPrime (425 genes) panel sequencing of the primary tumor, plasma and matched white blood cell samples from 20 PTC with DM and 46 PTC without DM. We identified somatic mutations, gene fusions and copy number alterations and analyzed their relationships with DM of PTC. BRAF-V600E was identified in 73% of PTC, followed by RET fusions (14%) in a mutually exclusive manner (P < 0.0001). We found that gene fusions (RET, ALK or NTRK1) (P < 0.01) and chromosome 22q loss (P < 0.01) were independently associated with DM in both univariate and multivariate analyses. A nomogram model consisting of chromosome 22q loss, gene fusions and three clinical variables was built for predicting DM in PTC (C-index = 0.89). The plasma circulating tumor DNA (ctDNA) detection rate in PTC was only 38.9%; however, it was significantly associated with the metastatic status (P = 0.04), tumor size (P = 0.001) and invasiveness (P = 0.01). In conclusion, gene fusions and chromosome 22q loss were independently associated with DM in PTC and could serve as molecular biomarkers for predicting DM. The ctDNA detection rate was low in non-DM PTC but significantly higher in PTC with DM.


Asunto(s)
Biomarcadores de Tumor/genética , Invasividad Neoplásica/genética , Cáncer Papilar Tiroideo/genética , Cáncer Papilar Tiroideo/patología , Neoplasias de la Tiroides/genética , Neoplasias de la Tiroides/patología , Adolescente , Adulto , Anciano , Niño , Femenino , Fusión Génica , Reordenamiento Génico , Humanos , Masculino , Persona de Mediana Edad , Mutación , Estudios Retrospectivos , Secuenciación del Exoma , Adulto Joven
8.
Appl Opt ; 59(28): 8848-8855, 2020 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-33104570

RESUMEN

It is difficult to obtain a large amount of labeled data, which has become a bottleneck for the application of deep learning to analyze one-dimensional optical time series signals. In order to solve this problem, a deep convolutional generative adversarial network model suitable for augmenting optical time series signals is proposed. Based on the acoustic emission (AE) data set obtained by an optical sensor with a small amount, the model can learn the corresponding data features and apply them to generate new data. The analysis results show that our model can generate stable and diverse AE fragments in epoch 500, and there is no model collapse. All the features between the generated data and the original data are not significantly different at the 0.05 level, which confirms that the method in this paper can generate the optical time series signals effectively.

9.
Curr Microbiol ; 77(3): 491-499, 2020 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-31832840

RESUMEN

Bacillus sp. TYF-LIM-B05, which is isolated from spoilage vinegar, is resistant to high temperature, high concentrated alcohol, acid, and salt, and can produce ethanol from mono-, di-, polysaccharide, and complex biomass as the sole carbon source. Thus, this strain is a potential candidate for consolidated bioprocessing (CBP) of fermenting lignocellulose to ethanol in a single step. To provide insight into the key enzymes involved in lignocellulose degradation and ethanol production, a draft genome of TYF-LIM-B05 was developed in this study. The results indicated that 348 genes are related to carbohydrate transport and metabolism according to the clusters of orthologous groups of proteins and annotated 187 CAZy domains from a total of 61 different families. The presence of genes encoding laccases, quinone oxidoreductases/reductases, and aryl-alcohol dehydrogenases further implies that TYF-LIM-B05 has the potential to degrade lignin. Remarkably, this strain has the ability to catalyze the oxidative decarboxylation of pyruvate to acetyl-CoA by pyruvate dehydrogenase complexes. The genomic information provided in this study will help researchers to better understand the mechanisms of the lignocellulose degradation and ethanol production pathway in thermophiles.


Asunto(s)
Bacillus/genética , Bacillus/metabolismo , Carbono/metabolismo , Etanol/metabolismo , Genoma Bacteriano , Lignina/metabolismo , Ácido Acético , Biomasa , Biología Computacional , Fermentación , Filogenia
10.
Opt Express ; 27(20): 28123-28132, 2019 Sep 30.
Artículo en Inglés | MEDLINE | ID: mdl-31684570

RESUMEN

This paper reports a torsion sensor based on the multimode interference theory. The sensor is fabricated by sandwiching a section of perfluorinated polymer optical fiber (POF) between two silica single mode fibers to construct a single-mode-multimode-single-mode (SMS) structure. The perfluorinated POF is easily connected to the optical fiber via the precise alignment of ceramic ferrules and ceramic mating sleeve. With the considerable flexibility and deformability of the perfluorinated POF, the proposed sensor is especially suitable for torsion measurement. Experimental results show that a wavelength sensitivity of 106.762 pm/(rad/m) and an intensity sensitivity of 0.165 dBm/(rad/m) are obtained within a large torsion rate of -100∼100 rad/m.

11.
Indoor Air ; 29(2): 215-230, 2019 03.
Artículo en Inglés | MEDLINE | ID: mdl-30474277

RESUMEN

Thermal comfort, self-reported acute health symptoms, cognitive performance, and physiological reactions were examined at four temperatures (26, 30, 33, and 37°C) at a relative humidity of 70%. Thirty-two sub-tropically acclimatized subjects experienced each condition for 175 minute, in balanced order, in a climatic chamber. The perception of heat gradually increased with increasing temperature, but the subjects felt hot only at 37°C. The temperature of 33°C was on average rated as acceptable and only just uncomfortable. The acceptability of air quality decreased linearly with increasing temperature. The intensity of acute health symptoms reported by the subjects increased with increasing temperature, but it was no more than moderate even at the highest temperature; dryness of skin and eye were alleviated. The eardrum temperature, skin temperature and moisture, heart rate, end-tidal carbon dioxide, and weight loss increased significantly with increasing temperature, whereas the percentage of adjacent heart inter-beat intervals differing by >50 ms decreased significantly. These results suggest that the perceived heat, self-reported symptoms, and physiological reactions occurred concurrently. They show additionally that acclimatization to heat may shift the boundary of thermal discomfort to a higher temperature. The role of psychological adaptation and of the contextual aspects of this process still requires clarification in future experiments.


Asunto(s)
Temperatura Corporal/fisiología , Estado de Salud , Calor/efectos adversos , Humedad , Sensación Térmica , Aclimatación , Adulto , Contaminación del Aire Interior , Análisis de Varianza , Peso Corporal , China , Cognición , Femenino , Humanos , Masculino , Temperatura Cutánea , Estudiantes , Encuestas y Cuestionarios , Temperatura , Universidades , Adulto Joven
12.
Appl Opt ; 58(26): 7251-7257, 2019 Sep 10.
Artículo en Inglés | MEDLINE | ID: mdl-31504001

RESUMEN

In this paper, a structural damage identification algorithm based on a single fiber Bragg grating (FBG) sensor is proposed. The signal detected by the FBG can be analyzed by the wavelet packet decomposition and back propagation neural network to obtain the damage location information. A high-speed FBG demodulation system based on a tunable Fabry-Perot filter and unbalanced Mach-Zehnder (M-Z) interferometer is designed to respond to a signal with a frequency range from 0 to 4 kHz, which will increase the sensing accuracy. This algorithm is verified by the aluminum plate model, which can simulate the generation of damage in reality. The experimental results show that a single FBG sensor is enough to realize accurate damage location identification according to this algorithm, and identification accuracy can reach 90.0%.

13.
Opt Express ; 26(18): 22944-22953, 2018 Sep 03.
Artículo en Inglés | MEDLINE | ID: mdl-30184951

RESUMEN

The thermal cycling process experienced by spacecraft during orbital operation would lead to deterioration of the demodulation performance of fiber Bragg grating (FBG). A new demodulation method based on Fabry-Perot (F-P) filter and hydrogen cyanide (HCN) gas is proposed to improve the performance. The method skillfully utilizes the self-marked HCN absorption lines as absolute wavelength references. In the thermal cycling environment whose temperature ranging from 5°Cto 65°C,the fluctuation of demodulation wavelength reduces to ± 2.6 pm, which is improved by 3.1 times compared with traditional method. The proposed method also shows a good robustness in the cases of weak light source intensity and poor signal-to-noise ratio (SNR) of HCN spectrum.

14.
Biotechnol Lett ; 40(2): 349-358, 2018 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-29124518

RESUMEN

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.


Asunto(s)
Oxidorreductasas de Alcohol/metabolismo , Amino Alcoholes/química , Amino Alcoholes/metabolismo , Biotecnología/métodos , Amino Alcoholes/análisis , Proteínas Bacterianas/metabolismo , Biocatálisis , Sistema Libre de Células , Escherichia coli/enzimología , Estereoisomerismo , Transaminasas/metabolismo
15.
Anal Biochem ; 518: 94-101, 2017 Feb 01.
Artículo en Inglés | MEDLINE | ID: mdl-27899283

RESUMEN

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.


Asunto(s)
Amino Alcoholes/química , Proteínas Bacterianas/química , Propanolaminas/química , Pseudomonas putida/enzimología , Transaminasas/química , Amino Alcoholes/metabolismo , Proteínas Bacterianas/metabolismo , Propanolaminas/metabolismo , Especificidad por Sustrato/fisiología , Transaminasas/metabolismo
16.
Biotechnol Lett ; 38(9): 1559-64, 2016 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-27233513

RESUMEN

OBJECTIVES: To investigate the efficiency of a cofactor regeneration enzyme co-expressed with a glycerol dehydrogenase for the production of 1,3-dihydroxyacetone (DHA). RESULTS: In vitro biotransformation of glycerol was achieved with the cell-free extracts containing recombinant GlyDH (glycerol dehydrogenase from Escherichia coli), LDH (lactate dehydrogenase form Bacillus subtilis) or LpNox1 (NADH oxidase from Lactobacillus pentosus), giving DHA at 1.3 g l(-1) (GlyDH/LDH) and 2.2 g l(-1) (GlyDH/LpNox1) with total turnover number (TTN) of NAD(+) recycling of 6039 and 11100, respectively. Whole cells of E. coli (GlyDH-LpNox1) co-expressing both GlyDH and LpNox1 were constructed and converted 10 g glycerol l(-1) to DHA at 0.2-0.5 g l(-1) in the presence of zero to 2 mM exogenous NAD(+). The cell free extract of E. coli (GlyDH-LpNox) converted glycerol (2-50 g l(-1)) to DHA from 0.5 to 4.0 g l(-1) (8-25 % conversion) without exogenous NAD(+). CONCLUSIONS: The disadvantage of the expensive consumption of NAD(+) for the production of DHA has been overcome.


Asunto(s)
Dihidroxiacetona/metabolismo , Glicerol/metabolismo , Complejos Multienzimáticos/metabolismo , NADH NADPH Oxidorreductasas/metabolismo , Deshidrogenasas del Alcohol de Azúcar/metabolismo
17.
Bioprocess Biosyst Eng ; 39(4): 603-11, 2016 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-26801669

RESUMEN

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.


Asunto(s)
Proteínas Bacterianas , Lactobacillus pentosus , NADPH Oxidasas , NAD/metabolismo , Proteínas Bacterianas/biosíntesis , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Catálisis , Clonación Molecular , Concentración de Iones de Hidrógeno , Lactobacillus pentosus/enzimología , Lactobacillus pentosus/genética , NAD/genética , NADPH Oxidasas/biosíntesis , NADPH Oxidasas/química , NADPH Oxidasas/genética , Oxidación-Reducción
18.
Biochemistry (Mosc) ; 80(2): 242-50, 2015 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-25756539

RESUMEN

Insect chitinase plays essential roles in chitin catabolism involved in digestion and molting during insect development. In the current work, we cloned a chitinase cDNA, LrCht5, from the apple leaf miner moth Lithocolletis ringoniella and characterized its amino acid sequence and protein properties. The L. ringoniella chitinase cDNA was 2136 bp in length with an open reading frame of 1737 bp that encodes a polypeptide of 579 amino acid residues with a predicted molecular mass of 64.4 kDa and pI of 5.49. The catalytic domain has several phosphorylation and glycosylation sites. The recombinant LrCht5 was expressed in Escherichia coli and the Spodoptera frugiperda cell line Sf9, and the LrCht5 expressed in insect cells exhibited chitinolytic activity. LrCht5 was most stable at pH 6.0 and 45°C. This work has potential application in the development of novel and more specific synthetic chitinase inhibitors for use as bioinsecticides.


Asunto(s)
Quitinasas/química , Mariposas Nocturnas/genética , Secuencia de Aminoácidos , Animales , Dominio Catalítico , Quitinasas/genética , Quitinasas/metabolismo , Clonación Molecular , Glicosilación , Datos de Secuencia Molecular , Fosforilación , Filogenia , Conformación Proteica
19.
Am J Transl Res ; 16(4): 1177-1187, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38715802

RESUMEN

OBJECTIVE: To investigate the impact of combining metformin with insulin aspart on blood glucose control, renal injury, and pregnancy outcome in gestational diabetes mellitus (GDM) patients. METHODS: In this retrospective analysis, the clinical data of 140 GDM patients treated at Baoji Maternal and Child Health Hospital between March 2020 and March 2022 were studied. The patients were divided into a control group (insulin aspart alone, n=64) and an observation group (combination of insulin aspart and metformin, n=76) according to their treatment regimen. The blood glucose metabolism, renal injury markers, and pregnancy outcomes between the two groups were assessed and compared. RESULTS: The observation group demonstrated significantly lower levels of blood glucose metabolism markers (fasting plasma glucose [FPG], fasting insulin [FINS], mean amplitude of glycemic excursions [MAGE], and mean of daily differences [MODD]), renal injury indicators (microalbuminuria [mAlb], serum cystatin C [CysC], free fatty acids [FFA], and neutrophil gelatinase-associated lipocalin [NGAL]), and inflammatory markers (interleukin-6 [IL-6], transforming growth factor-ß1 [TGF-ß1], and lipoprotein-associated phospholipase A2 [Lp-PLA2]) compared to the control group (all P<0.05). Additionally, the incidence of adverse pregnancy outcomes in both newborns and mothers was lower in the observation group (P<0.05). Logistic regression analysis identified the treatment regimen, patient age, and pre-pregnancy BMI as independent risk factors for adverse pregnancy outcome. CONCLUSION: The combination of metformin and insulin aspart in treating GDM can effectively reduce blood glucose levels, mitigate renal injury, and improve pregnancy outcome. This treatment approach presents a viable option for optimizing maternal and fetal health in GDM cases.

20.
Am J Transl Res ; 16(4): 1199-1208, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38715823

RESUMEN

OBJECTIVE: To investigate the correlation between serum thyroid-stimulating hormone (TSH) and total bile acid (TBA) levels in gestational hypertension and their combined predictive value for pregnancy outcome. METHODS: A total of 194 patients with gestational hypertension (GH), treated from June 2020 to May 2022, were included in this study. The patients were divided into two subgroups based on pregnancy outcome: an adverse pregnancy outcome group (77 cases) and a normal pregnancy outcome group (117 cases). Additionally, 50 healthy pregnant women undergoing routine prenatal checkups during the same period served as the control group. In this study, serum TBA and TSH levels were measured and compared between the control and GH groups as well as between adverse pregnancy outcome and normal pregnancy outcome groups. The independent risk factors for adverse pregnancy outcome were screened using logistic regression, and their predictive value for pregnancy outcome in patients with GH was analyzed using receiver operating characteristic (ROC) curves. RESULTS: Serum TSH and TBA levels were significantly higher in the GH group compared to the normal group (both P < 0.001). Logistic regression analysis revealed that age, body mass index (BMI), TSH, and TBA were independent risk factors for adverse pregnancy outcome. ROC curve analysis showed that combined TSH and TBA for predicting adverse pregnancy outcome had an Area Under the Curve (AUC) of 0.896, surpassing the AUCs of each individual index (0.843 for TSH and 0.765 for TBA), which indicates a stronger predictive value (P < 0.001). CONCLUSION: The combined measurement of serum TBA and TSH can serve as a valuable predictive tool for pregnancy outcome in patients with gestational hypertension.

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