RESUMEN
ATP-dependent energy-consuming enzymatic reactions are widely used in cell-free biocatalysis. However, the direct addition of large amounts of expensive ATP can greatly increase cost, and enzymatic production is often difficult to achieve as a result. Although a polyphosphate kinase (PPK)-polyphosphate-based ATP regeneration system has the potential to solve this challenge, the generally poor thermal stability of PPKs limits the widespread use of this method. In this paper, we evaluated the thermal stability of a PPK from Sulfurovum lithotrophicum (SlPPK2). After directed evolution and computation-supported design, we found that SlPPK2 is very recalcitrant and cannot acquire beneficial mutations. Inspired by the usually outstanding stability of ancestral enzymes, we reconstructed the ancestral sequence of the PPK family and used it as a guide to construct three heat-stable variants of SlPPK2, of which the L35F/T144S variant has a half-life of more than 14 h at 60°C. Molecular dynamics simulations were performed on all enzymes to analyze the reasons for the increased thermal stability. The results showed that mutations at these two positions act synergistically from the interior and surface of the protein, leading to a more compact structure. Finally, the robustness of the L35F/T144S variant was verified in the synthesis of nucleotides at high temperature. In practice, the use of this high-temperature ATP regeneration system can effectively avoid byproduct accumulation. Our work extends the temperature boundary of ATP regeneration and has great potential for industrial applications.IMPORTANCEATP regeneration is an important basic applied study in the field of cell-free biocatalysis. Polyphosphate kinase (PPK) is an enzyme tool widely used for energy regeneration during enzymatic reactions. However, the thermal stability of the PPKs reported to date that can efficiently regenerate ATP is usually poor, which greatly limits their application. In this study, the thermal stability of a difficult-to-engineer PPK from Sulfurovum lithotrophicum was improved, guided by an ancestral sequence reconstruction strategy. The optimal variant has a 4.5-fold longer half-life at 60°C than the wild-type enzyme, thus enabling the extension of the temperature boundary for ATP regeneration. The ability of this variant to regenerate ATP was well demonstrated during high-temperature enzymatic production of nucleotides.
Asunto(s)
Adenosina Trifosfato , Epsilonproteobacteria , Fosfotransferasas (Aceptor del Grupo Fosfato) , Adenosina Trifosfato/metabolismo , Temperatura , Fosfotransferasas (Aceptor del Grupo Fosfato)/genética , Fosfotransferasas (Aceptor del Grupo Fosfato)/metabolismo , NucleótidosRESUMEN
PURPOSE: The optimal surgical procedure, whether total gastrectomy (TG) or proximal gastrectomy (PG), for Siewert type II/III adenocarcinoma of esophagogastric junction (AEG) has not been standardised, primarily because the optimal extent of lymph node (LN) dissection for AEG based on the metastatic rate of perigastric LNs remains under debate. The aim of this study was to investigate the metastatic incidence and prognostic significance of lower perigastric lymph nodes (LPLN), including No.4d, 5, 6 and 12a LN stations, in Siewert type II/III AEG. METHODS: A total of 701 patients with Siewert type II/III AEG who received transabdominal open gastrectomy (425 patients with TG and 276 patients with PG) from 2010 to 2015 in West China Hospital were retrospectively included. Based on the clinicopathological information of TG patients, the risk factors of LPLN-positive patients were evaluated, and the metastatic incidence as well as the therapeutic value (TV) index of each LN station was assessed. Moreover, the 5-year overall survival (OS) rates between LPLN-positive and LPLN-negative groups were compared in TG patients, and the postoperative survival difference between TG and PG patients was also compared, using propensity score matching (PSM) method. RESULTS: Tumour size (≥ 5 cm, OR = 1.481, p = 0.002) and pT stage (pT4, OR = 2.755, p = 0.024) were significant risk factors for patients with LPLN metastasis. For patients with tumour size more than 5 cm or pT4 stage, the metastatic rates of LPLN for Siewert type II, III and II/III AEG were 31.67%, 34.69% and 33.03%, whereas the TV indexes of LPLN for them were 5.76, 5.62 and 5.38, respectively. LPLN was a significant independent prognostic factor (HR = 1.422, p = 0.028), and positive LPLN was related to worse prognosis (p < 0.05). For patients with tumour size more than 5 cm or pT4 stage, TG patients were illustrated to have a better prognosis than PG patients, with 5-year OS rates of 58.9% vs 38.2% for Siewert type II AEG (χ2 = 4.159, p = 0.041), 68.9% vs 50.2% for Siewert type III AEG (χ2 = 5.630, p = 0.018) and 65.1% vs 40.3% for Siewert type II/III AEG (χ2 = 12.604, p < 0.001), respectively. CONCLUSIONS: LPLN metastasis is a poor prognostic factor for patients with Siewert II/III AEG. LPLN dissection may improve the long-term survival of patients with tumour size more than 5 cm or pT4 stage, and TG might be more suitable for this kind of cancer.
Asunto(s)
Adenocarcinoma , Neoplasias Esofágicas , Neoplasias Gástricas , Adenocarcinoma/patología , Neoplasias Esofágicas/patología , Neoplasias Esofágicas/cirugía , Unión Esofagogástrica/patología , Unión Esofagogástrica/cirugía , Gastrectomía/métodos , Humanos , Escisión del Ganglio Linfático/métodos , Ganglios Linfáticos/patología , Metástasis Linfática/patología , Puntaje de Propensión , Estudios Retrospectivos , Neoplasias Gástricas/patologíaRESUMEN
We investigate threshold current temperature dependence of electrically injected quantum-dot (QD) photonic crystal (PC) surface-emitting lasers (SELs) with respect to wavelength detuning between QD gain peak and PC cavity resonance. The lasing emissions cover wavelengths from 1283 nm to 1318 nm. Almost infinite characteristic temperature is realized at certain temperature range for PCSEL with large negative gain-cavity detuning. Moreover, band-edge lasing mode is identified in our "PC slab-on-substrate" structure, and its far-field distribution is characterized as doughnut-shaped beam with azimuthal polarization.
RESUMEN
Recently, the moiré pattern has attracted lots of attention by superimposing two planar structures of regular geometries, such as two sets of metasurfaces or gratings. Here, we show the experimental investigation of acoustic moiré effect by using twisted bilayer gratings (i.e., one grating twisted with respect to the other). We observed the guided resonance that occurred when the incident ultrasound beam was coupled with the guiding modes in a meta-grating, significantly influencing the reflection and transmission. Tunable guided resonances from the moiré effect with complete ultrasound reflection at different frequencies were further demonstrated in experiments. Combining the measurements of transmission spectra and the Fast Fourier Transform analyses, we reveal the guided resonance frequencies of moiré ultrasonic metasurface can be effectively controlled by adjusting the twisting angle of the bilayer gratings. Our results can be explained in a simplified model based on the band folding theory, providing a reliable prediction on the precise control of ultrasound reflection via the twisting angle adjustment. Our work extends the moiré metasurface from optics into acoustics, which shows more possibilities for the ultrasound beam engineering from the moiré effect and enables the exploration of functional acoustic devices for ultrasound imaging, treatment and diagnosis.
RESUMEN
Objective: Recent studies have indicated potential anti-inflammatory effects of glucagon-like peptide-1 receptor agonists (GLP-1RAs) on asthma, which is often comorbid with type 2 diabetes mellitus (T2DM) and obesity. Therefore, we conducted a meta-analysis to assess the association between the administration of glucagon-like peptide-1 (GLP-1) receptor-based agonists and the incidence of asthma in patients with T2DM and/or obesity. Methods: PubMed, Web of Science, Embase, the Cochrane Central Register of Controlled Trials, and Clinicaltrial.gov were systematically searched from inception to July 2023. Randomized controlled trials (RCTs) of GLP-1 receptor-based agonists (GLP-1RA, GLP-1 based dual and triple receptor agonist) with reports of asthma events were included. Outcomes were computed as risk ratios ( RR) using a fixed-effects model. Results: Overall, 39 RCTs with a total of 85,755 participants were included. Compared to non-GLP-1 receptor-based agonist users, a trend of reduced risk of asthma was observed in patients with T2DM or obesity using GLP-1 receptor-based agonist treatments, although the difference was not statistically significant [ RR = 0.91, 95% confidence interval ( CI): 0.68 to 1.24]. Further Subgroup analyses indicated that the use of light-molecular-weight GLP-1RAs might be associated with a reduced the risk of asthma when compared with non-users ( RR = 0.65, 95% CI: 0.43 to 0.99, P = 0.043). We also performed sensitivity analyses for participant characteristics, study design, drug structure, duration of action, and drug subtypes. However, no significant associations were observed. Conclusion: Compared with non-users, a modest reduction in the incidence of asthma was observed in patients with T2DM or obesity using GLP-1 receptor-based agonist treatments. Further investigations are warranted to assess the association between GLP-1 receptor-based agonists and the risk of asthma.
Asunto(s)
Asma , Diabetes Mellitus Tipo 2 , Receptor del Péptido 1 Similar al Glucagón , Obesidad , Humanos , Asma/epidemiología , Asma/prevención & control , Diabetes Mellitus Tipo 2/complicaciones , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Receptor del Péptido 1 Similar al Glucagón/agonistas , Hipoglucemiantes/uso terapéutico , Incidencia , Obesidad/complicacionesRESUMEN
We have proposed and fabricated a new mid-infrared reflector using the guided-mode resonance (GMR). The GMR reflector consists of subwavelength Ge grating on GaAs substrate with a low-refractive-index SiOx layer in between. With a total thickness of about 2 µm, a near-100% reflectivity at 8 µm has been obtained both theoretically and experimentally.
RESUMEN
BACKGROUND: Chiglitazar is an emerging pan-agonist of all peroxisome proliferator activated receptors (PPAR)-α, δ and γ, and has therapeutic potential for type 2 diabetes (T2D). However, to date, no clinical studies or meta-analyses have compared the efficacy and safety of chiglitazar and traditional PPAR-γ agonist thiazolidinediones (TZDs). A meta-analysis concerning this topic is therefore required. AIM: To compare the efficacy and safety of chiglitazar and TZD in patients with T2D. METHODS: PubMed, Medline, Embase, the Cochrane Central Register of Controlled Trials, Reference Citation Analysis and Clinicaltrial.gov websites were searched from August 1994 to March 2022. Randomized controlled trials (RCTs) of chiglitazar or TZD vs placebo in patients with T2D were included. Indirect comparisons and sensitivity analyses were implemented to evaluate multiple efficacy and safety endpoints of interest. RESULTS: We included 93 RCTs that compared TZD with placebo and one that compared chiglitazar with placebo. For efficacy endpoints, the augmented dose of chig-litazar resulted in greater reductions in hemoglobin (Hb)A1c [weighted mean difference (WMD) = -0.15%, 95% confidence interval (CI): -0.27 to -0.04%], triglycerides (WMD = -0.17 mmol/L, 95%CI: -0.24 to -0.11 mmol/L) and alanine aminotransferase (WMD = -5.25 U/L, 95%CI: -8.50 to -1.99 U/L), and a greater increase in homeostasis model assessment-ß (HOMA-ß) (WMD = 17.75, 95%CI: 10.73-24.77) when compared with TZD treatment. For safety endpoints, the risks of hypoglycemia, edema, bone fractures, upper respiratory tract infection, urinary tract infection, and weight gain were all comparable between the augmented dose of chiglitazar and TZD. In patients with baseline HbA1c ≥ 8.5%, body mass index ≥ 30 kg/m2 or diabetes duration < 10 years, the HbA1c reduction and HOMA-ß increase were more conspicuous for the augmented dose of chiglitazar compared with TZD. CONCLUSION: Augmented dose of chiglitazar, a pan-activator of PPARs, may serve as an antidiabetic agent with preferable glycemic and lipid control, better ß-cell function preserving capacity, and does not increase the risk of safety concerns when compared with TZD.
RESUMEN
Cellulose, as a component of green plants, becomes attractive for fabricating biocompatible flexible functional devices but is plagued by hydrophilic properties, which make it easily break down in water by poor mechanical stability. Here we report a class of SiO2-nanoparticle-decorated bacteria-cellulose meta-skin with superior stability in water, excellent machining property, ultrathin thickness, and active bacteria-repairing capacity. We further develop functional ultrasonic metasurfaces based on meta-skin paper-cutting that can generate intricate patterns of ~10 µm precision. Benefited from the perfect ultrasound insulation of surface Cassie-Baxter states, we utilize meta-skin paper-cutting to design and fabricate ultrathin (~20 µm) and super-light (<20 mg) chip-scale devices, such as nonlocal holographic meta-lens and the 3D imaging meta-lens, realizing complicated acoustic holograms and high-resolution 3D ultrasound imaging in far fields. The decorated bacteria-cellulose ultrasonic metasurface opens the way for exploiting flexible and biologically degradable metamaterial devices with functionality customization and key applications in advanced biomedical engineering technologies.
RESUMEN
Hepatitis B e antigen (HBeAg) is a widely used marker both for chronic hepatitis B (CHB) clinical management and HBV-related basic research. However, due to its high amino acid sequence homology to hepatitis B core antigen (HBcAg), most of available anti-HBe antibodies are cross-reactive with HBcAg resulting in high interference against accurate measurement of the status and level of HBeAg. In the study, we generated several monoclonal antibodies (mAbs) targeting various epitopes on HBeAg and HBcAg. Among these mAbs, a novel mAb 16D9, which recognizes the SKLCLG (aa -10 to -5) motif on the N-terminal residues of HBeAg that is absent on HBcAg, exhibited excellent detection sensitivity and specificity in pairing with another 14A7 mAb targeting the HBeAg C-terminus (STLPETTVVRRRGR, aa141 to 154). Based on these two mAbs, we developed a novel chemiluminescent HBeAg immunoassay (NTR-HBeAg) which could detect HBeAg derived from various HBV genotypes. In contrast to widely used commercial assays, the NTR-HBeAg completely eliminated the cross-reactivity with secreted HBcAg from precore mutant (G1896A) virus in either cell culture or patient sera. The improved specificity of the NTR-HBeAg assay enables its applicability in cccDNA-targeting drug screening in cell culture systems and also provides an accurate tool for clinical HBeAg detection.
Asunto(s)
Anticuerpos contra la Hepatitis B/análisis , Antígenos e de la Hepatitis B/química , Virus de la Hepatitis B/genética , Hepatitis B Crónica/inmunología , Secuencias de Aminoácidos , Anticuerpos Monoclonales/análisis , Técnicas de Cultivo de Célula , Línea Celular , Epítopos/inmunología , Genotipo , Células Hep G2 , Antígenos del Núcleo de la Hepatitis B/química , Antígenos del Núcleo de la Hepatitis B/inmunología , Antígenos e de la Hepatitis B/inmunología , Virus de la Hepatitis B/inmunología , Hepatitis B Crónica/sangre , Humanos , Mediciones LuminiscentesRESUMEN
Coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), was found initially in Wuhan, China in early December 2019. The pandemic has spread to 216 countries and regions, infecting more than 23310 000 people and causing over 800 000 deaths globally by Aug. 24, 2020, according to World Health Organization (https://www.who.int/emergencies/diseases/ novel-coronavirus-2019). Fever, cough, and dyspnea are the three common symptoms of the condition, whereas the conventional transmission route for SARS-CoV-2 is through droplets entering the respiratory tract. To date, infection control measures for COVID-19 have been focusing on the involvement of the respiratory system. However, ignoring potential faecal transmission and the gastrointestinal involvement of SARS-CoV-2 may result in mistakes in attempts to control the pandemic.
Asunto(s)
Betacoronavirus/aislamiento & purificación , Infecciones por Coronavirus/transmisión , Infecciones por Coronavirus/virología , Heces/virología , Enfermedades Gastrointestinales/virología , Neumonía Viral/transmisión , Neumonía Viral/virología , Betacoronavirus/genética , COVID-19 , China/epidemiología , Infecciones por Coronavirus/epidemiología , Microbiología Ambiental , Humanos , Modelos Biológicos , Pandemias , Neumonía Viral/epidemiología , ARN Viral/análisis , ARN Viral/genética , SARS-CoV-2 , Esparcimiento de VirusRESUMEN
We study the effect of etching depth on the threshold characteristics of GaSb-based middle infrared (Mid-IR) photonic-crystal surface-emitting lasers (PCSELs) with different lattice periods. The below-threshold emission spectra are measured to identify the bandgap as well as band-edge modes. Moreover, the bandgap separation widens with increasing etching depth as a result of enhanced diffraction feedback coupling. However, the coupling is nearly independent of lattice period. The relationship between threshold gain and Bragg detuning is also experimentally determined for PCSELs and is similar to that calculated theoretically for one-dimensional distributed feedback lasers.
RESUMEN
This paper discusses the issue of controlling the epitaxial growth of mixed group V alloys to form a type-I InGaAsSb/AlGaAsSb double quantum wells (QWs) structure. We also discuss the run-to-run reproducibility of lattice-matched AlGaAsSb alloys and strained In0.35Ga0.65As0.095Sb0.905 in terms of growth parameters (V/III ratio, Sb2/As2 ratio). Molecular beam epitaxy (MBE) was used to grow two type-I InGaAsSb double-QWs laser structures differing only in the composition of the bottom cladding layer: Al0.85Ga0.15As0.072Sb0.928 (sample A) and Al0.5Ga0.5As0.043Sb0.957 (sample B). Both samples were respectively used in the fabrication of photonic crystal surface-emitting lasers (PCSELs). Sample A presented surface lasing action from circular as well as triangular photonic crystals. Sample B did not present surface lasing due to the deterioration of the active region during the growth of the upper cladding. Our findings underline the importance of temperature in the epitaxial formation of AlxGa1-xAsySb1-y in terms of lasing performance.
RESUMEN
Highly branched metallic nanostructures, which possess a large amount of catalyst active sites and surface-enhanced Raman scattering (SERS) hot spots owing to their large surface areas, multi-level branches, corners, and edges, have shown potential in various applications including catalysis and SERS. In this study, well-defined dendritic silver (Ag) nanostructures were prepared by a facile and controllable electrochemical deposition strategy. The morphology of Ag nanostructures is controlled by regulating electrodeposition time and concentration of AgNO3 in the electrolyte solution. Compared to conventional Ag nanoparticle films, dendritic Ag nanostructures exhibited larger SERS enhancement ascribed to the numerous hot spots exist in the nanogaps of parallel and vertically stacked multilayer Ag dendrites. In addition, the prepared dendritic Ag nanostructures show 3.2-fold higher catalytic activity towards the reduction of 4-nitrophenol (4-NP) by NaBH4 than the Ag nanoparticle films. The results indicate that the dendritic Ag nanostructures represent a unique bifunctional nanostructure that serves as both efficient catalysts and excellent SERS substrates, which may be further employed as a nanoreactor for in situ investigation and real-time monitoring of catalytic reactions by SERS technique.
RESUMEN
Increasing evidence suggests that microRNAs (miRNAs) play a critical role in tumorigenesis. Decreased expression of miR-382 has been observed in various types of cancers. However, the biological function of miR-382 in colorectal cancer (CRC) is still largely unknown. Here, we found that miR-382 was down-regulated in human colorectal cancer tissues and cell lines associated with it. MiR-382 inhibited colorectal cancer cell proliferation, migration, invasion, and enhance chemosensitivity. Furthermore, we identified Krüppel-like factor 12 (KLF12) and homeodomain-interacting protein kinase 3 (HIPK3) as the target of miR-382, and miR-382 rescued the promotion effect of KFL12 on migration and enhanced chemosensitivity in colorectal cancer cell lines. Collectively, these findings revealed that miR-382 inhibits migration and enhances chemosensitivity by targeting KLF12 and HIPK3 in colorectal cancer. These findings might serve as a tumor suppressor in CRC.
Asunto(s)
Neoplasias Colorrectales/genética , Genes Supresores de Tumor/fisiología , MicroARNs/genética , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular/genética , Regulación hacia Abajo/genética , Regulación Neoplásica de la Expresión Génica/genética , Células HCT116 , Humanos , Factores de Transcripción de Tipo Kruppel/genéticaRESUMEN
OBJECTIVE: To explore the feasibility and clinical value of secondary sentinel lymph node (SSLN) tracing technique in radical gastrectomy for advanced gastric cancer (AGC). METHODS: From January 2009 to June 2011, 247 patients who suffered from gastric angle cancer with metastasis in No. 3 group lymph nodes were divided randomly into groups A and B. Methylthioninium chloride was injected into the peripheral tissue of the metastatic No. 3 group lymph nodes of 138 patients in group A before tumor resections. SSLNs were traced and individual lymphadenectomies were carried out based on the biopsy results of the SSLNs. Standard D2 radical gastrectomies were carried out directly on 109 patients in group B. Postoperative follow-up and survival analysis were carried out for patients in both groups. RESULTS: SSLNs were found in 114 (82.6%) patients in group A. Ninety of those patients (78.9%) demonstrated existing metastasis in SSLNs. According to Kaplan-Meier's method, the postoperative 3-year cumulative survival rates were 63.5% and 47.5%, and the median survival time were 40 and 36 months for the patients of groups A and B, respectively (P<0.05). CONCLUSIONS: The SSLN tracing technique is feasible in radical gastrectomy for AGC. It gives surgeons important information about the terminal status of lymph node metastasis and provides some scientific basis for individual lymphadenectomy.
Asunto(s)
Gastrectomía , Escisión del Ganglio Linfático , Ganglios Linfáticos/patología , Neoplasias Gástricas/patología , Adulto , Anciano , Femenino , Humanos , Escisión del Ganglio Linfático/métodos , Metástasis Linfática , Masculino , Persona de Mediana Edad , Biopsia del Ganglio Linfático Centinela , Neoplasias Gástricas/cirugíaRESUMEN
OBJECTIVES: To study the inhibitory effect of KF-950 on human acrosin and sperm acrosome. METHODS: Human acrosin was extracted and purified with 2% acetic acid, and its residual activity was evaluated by BAEE/ADH assay after treated with different concentrations of KF-950. ABC assay was used to observe the effect of KF-950 on human acrosome with Biotin-PSA as a probe. RESULTS: 1. The activity of normal sperm acrosin was (37.65 +/- 4.47) U/L. 2. The residual activity was inversely related to the concentration of KF-950 (r = -0.998), and had a dose-response curve. The result could be described by Y = 7.57-1.895X. 3. With increase of KF-950 concentration and prolongation of action time, the staining rate of acrosome obviously dropped (P < 0.01). CONCLUSIONS: KF-950 directly inhibits acrosin activity and assumely injures sperm acrosome. It might be a new kind of highly effective inhibitor.
Asunto(s)
Acrosina/antagonistas & inhibidores , Acrosoma/metabolismo , Espermatozoides/efectos de los fármacos , Humanos , Masculino , Espermatozoides/fisiologíaRESUMEN
OBJECTIVE: To study dynamic relation between periodontal pathogens and cariogenic bacteria under analogous oral environment. METHODS: Eight periodontopathic and cariogenic bacteria of Porphyromonas gingivalis (Pg), Actinobacillus actinomycetemcomitans (Aa), Fusobacterium nucleatum (Fn), Provotella intermedium (Pi), Streptococcus mutans (Sm), Streptococcus sanguis (Ss), Actinomyces viscosus (Av) and Lactobacillus acidophilus (La) were used. These eight strains were cultured in modified chemostat under analogous oral environment which contained 600 ml modified BM medium supplemented with 2.5 g/L porcine gastric mucin, respectively. After 1, 24, 48 and 96 h, optical sectioning of plaque biofilms on removable and replaceable hydroxyapatite disks was analyzed by the combination of live bacterial Gram fluorescence staining and confocal laser scanning microscopy. Biofilm thickness and reconstruction of the three-dimensional architecture of plaque biofilms were made. RESULTS: Biofilm thickness increased significantly with time (P < 0.001). Biofilms of Aa were thinner than those of Ss and eight-specie biofilms were thicker than those formed by Ss and Aa per time point. Three-dimensional images showed periodontal pathogens mainly occurred in cariogenic bacterial complex or on the biofilm surface. CONCLUSIONS: Gram-positive cariogenic species initially predominated in artificial plaque, followed by the increasing proportions of Gram-negative periodontal pathogens. The relation between microecological balance among bacteria and diseases is worthy of further studies.