Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 24.030
Filtrar
1.
Spectrochim Acta A Mol Biomol Spectrosc ; 324: 124914, 2025 Jan 05.
Artículo en Inglés | MEDLINE | ID: mdl-39137711

RESUMEN

Lead (Pb) is a heavy metal known for its adverse effects on both human health and the environment. In recent years, the industrial utilization of Pb2+ has surged, underscoring the imperative need for efficient measurement methods. In this study, a rapid and simple photochemical method was used to synthesize thioglycolic acid (TGA)-stabilized CdTe/ZnSe core-shell quantum dots (QDs). These CdTe/ZnSe QDs emit vibrant green fluorescence and exhibit remarkable quenching in the presence of Pb2+ ions. This property enables the development of an on-site on/off sensor without the necessity of additional modifications. The proposed sensor possesses an outstanding sensitivity to Pb2+, with a detection limit and linear range of 31.8 nM and 50 nM-10 µM, respectively. Importantly, the selectivity of this fluorescence-based sensor was validated by analyzing various positively and negatively charged ions. Furthermore, the developed sensor showed reliable performance against real river, agricultural, and tap water, as confirmed by Inductively Coupled Plasma (ICP) analysis. Additionally, CdTe/ZnSe QDs immobilized on glass slides were successfully employed for on-site water sample analysis, providing a versatile solution for environmental monitoring.

2.
Food Chem ; 462: 141063, 2025 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-39226640

RESUMEN

In this research, the TT-COF(Fe)@NH2-CNTs was innovatively prepared through a post-modification synthetic process functionalized TT-COF@NH2-CNTs with active site (Fe), where TT-COF@NH2-CNTs was prepared via a one-pot strategy using 5,10,15,20-tetrakis (para-aminophenyl) porphyrin (TTAP), 2,3,6,7-tetra (4-formylphenyl) tetrathiafulvalene (TTF) and aminated carbon nanotubes (NH2-CNTs) as raw materials. The complex TT-COF(Fe)@NH2-CNTs material possessed porous structures, outstanding conductivity and rich catalytic sites. Thus, it can be adopted to construct electrochemical sensor with glassy carbon electrode (GCE). The TT-COF(Fe)@NH2-CNTs/GCE can selectively detect luteolin (Lu) with a wide linear plot ranging from 0.005 to 3 µM and a low limit of detection (LOD) of 1.45 nM (S/N = 3). The Lu residues in carrot samples were determined using TT-COF(Fe)@NH2-CNTs sensor and UV-visible (UV-Vis) approach. This TT-COF(Fe)@NH2-CNTs/GCE sensor paves the way for the quantification of Lu through a cost-efficient and sensitive electrochemical approach, which can make a significant step in the sensing field based on crystalline COFs.


Asunto(s)
Técnicas Electroquímicas , Luteolina , Nanotubos de Carbono , Nanotubos de Carbono/química , Luteolina/química , Luteolina/análisis , Técnicas Electroquímicas/instrumentación , Límite de Detección , Estructuras Metalorgánicas/química , Contaminación de Alimentos/análisis , Dominio Catalítico
3.
J Environ Sci (China) ; 148: 306-320, 2025 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-39095167

RESUMEN

Antimony smelting activities damage the soil and vegetation surroundings while generating economic value. However, no standardized methods are available to diagnose the extent of soil degradation at antimony smelting sites. This study developed a standardized framework for assessing soil quality by considering microbial-induced resilience and heavy metal contamination at Xikuangshan antimony smelting site. The soil resilience index (SRI) and soil contamination index (SCI) were calculated by Minimum Data Set and geo-accumulation model, respectively. After standardized by a multi-criteria quantitative procedure of modified Nemerow's pollution index (NPI), the integrated assessment of soil quality index (SQI), which is the minimum of SRINPI and SCINPI, was achieved. The results showed that Sb and As were the prominent metal(loid) pollutants, and significant correlations between SQI and SRI indicated that the poor soil quality was mainly caused by the low level of soil resilience. The primary limiting factors of SRI were Fungi in high and middle contaminated areas, and Skermanella in low contaminated area, suggesting that the weak soil resilience was caused by low specific microbial abundances. Microbial regulation and phytoremediation are greatly required to improve the soil quality at antimony smelting sites from the perspectives of pollution control and resilience improvement. This study improves our understanding of ecological effects of antimony smelting sites and provides a theoretical basis for ecological restoration and sustainable development of mining areas.


Asunto(s)
Antimonio , Monitoreo del Ambiente , Metales Pesados , Microbiología del Suelo , Contaminantes del Suelo , Suelo , Contaminantes del Suelo/análisis , Antimonio/análisis , Monitoreo del Ambiente/métodos , Metales Pesados/análisis , Suelo/química , Metalurgia , Biodegradación Ambiental , China
4.
J Environ Sci (China) ; 149: 242-253, 2025 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-39181639

RESUMEN

Poly(butylene succinate-co-furandicarboxylate) (PBSF) and poly(butylene adipate-co-furandicarboxylate) (PBAF) are novel furandicarboxylic acid-based biodegradable copolyesters with great potential to replace fossil-derived terephthalic acid-based copolyesters such as poly(butylene succinate-co-terephthalate) (PBST) and poly(butylene adipate-co-terephthalate) (PBAT). In this study, quantum chemistry techniques after molecular dynamics simulations are employed to investigate the degradation mechanism of PBSF and PBAF catalyzed by Candida antarctica lipase B (CALB). Computational analysis indicates that the catalytic reaction follows a four-step mechanism resembling the ping-pong bibi mechanism, with the initial two steps being acylation reactions and the subsequent two being hydrolysis reactions. Notably, the first step of the hydrolysis is identified as the rate-determining step. Moreover, by introducing single-point mutations to expand the substrate entrance tunnel, the catalytic distance of the first acylation step decreases. Additionally, energy barrier of the rate-determining step is decreased in the PBSF system by site-directed mutations on key residues increasing hydrophobicity of the enzyme's active site. This study unprecedently show the substrate binding pocket and hydrophobicity of the enzyme's active site have the potential to be engineered to enhance the degradation of copolyesters catalyzed by CALB.


Asunto(s)
Proteínas Fúngicas , Lipasa , Poliésteres , Lipasa/metabolismo , Lipasa/química , Proteínas Fúngicas/metabolismo , Proteínas Fúngicas/química , Poliésteres/química , Poliésteres/metabolismo , Biodegradación Ambiental , Simulación de Dinámica Molecular , Hidrólisis , Modelos Químicos
5.
Biomaterials ; 312: 122755, 2025 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-39151270

RESUMEN

Copper-catalyzed click chemistry offers creative strategies for activation of therapeutics without disrupting biological processes. Despite tremendous efforts, current copper catalysts face fundamental challenges in achieving high efficiency, atom economy, and tissue-specific selectivity. Herein, we develop a facile "mix-and-match synthetic strategy" to fabricate a biomimetic single-site copper-bipyridine-based cerium metal-organic framework (Cu/Ce-MOF@M) for efficient and tumor cell-specific bioorthogonal catalysis. This elegant methodology achieves isolated single-Cu-site within the MOF architecture, resulting in exceptionally high catalytic performance. Cu/Ce-MOF@M favors a 32.1-fold higher catalytic activity than the widely used MOF-supported copper nanoparticles at single-particle level, as first evidenced by single-molecule fluorescence microscopy. Furthermore, with cancer cell-membrane camouflage, Cu/Ce-MOF@M demonstrates preferential tropism for its parent cells. Simultaneously, the single-site CuII species within Cu/Ce-MOF@M are reduced by upregulated glutathione in cancerous cells to CuI for catalyzing the click reaction, enabling homotypic cancer cell-activated in situ drug synthesis. Additionally, Cu/Ce-MOF@M exhibits oxidase and peroxidase mimicking activities, further enhancing catalytic cancer therapy. This study guides the reasonable design of highly active heterogeneous transition-metal catalysts for targeted bioorthogonal reactions.


Asunto(s)
Materiales Biomiméticos , Cobre , Humanos , Cobre/química , Materiales Biomiméticos/química , Catálisis , Estructuras Metalorgánicas/química , Neoplasias/tratamiento farmacológico , Neoplasias/terapia , Cerio/química , Línea Celular Tumoral , Animales , Química Clic/métodos , Biomimética/métodos , Ratones
6.
ACS Synth Biol ; 13(9): 3022-3031, 2024 Sep 20.
Artículo en Inglés | MEDLINE | ID: mdl-39238421

RESUMEN

Genome integration enables host organisms to stably carry heterologous DNA messages, introducing new genotypes and phenotypes for expanded applications. While several genome integration approaches have been reported, a scalable tool for DNA message storage within site-specific genome landing pads is still lacking. Here, we introduce an iterative genome integration method utilizing orthogonal serine integrases, enabling the stable storage of multiple heterologous genes in the chromosome of Escherichia coli MG1655. By leveraging serine integrases TP901-1, Bxb1, and PhiC31, along with engineered integration vectors, we demonstrate high-efficiency, marker-free integration of DNA fragments up to 13 kb in length. To further simplify the procedure, we then develop a streamlined integration method and showcase the system's versatility by constructing an engineered E. coli strain capable of storing and expressing multiple genes from diverse species. Additionally, we illustrate the potential utility of these engineered strains for synthetic biology applications, including in vivo and in vitro protein expression. Our work extends the application scope of serine integrases for scalable gene integration cascades, with implications for genome manipulation and gene storage applications in synthetic biology.


Asunto(s)
Escherichia coli , Genoma Bacteriano , Integrasas , Escherichia coli/genética , Genoma Bacteriano/genética , Integrasas/genética , Integrasas/metabolismo , Biología Sintética/métodos , Serina/metabolismo , Serina/genética , Ingeniería Genética/métodos , Vectores Genéticos/genética
7.
Environ Sci Pollut Res Int ; 31(43): 55648-55662, 2024 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-39240437

RESUMEN

In the study, the structural parameters of Zichang (ZC) coking coal from northern Shaanxi Province were examined. A theoretical calculation was employed to build a molecular structure model for ZC coal, as well as applying principles of quantum chemistry, the prediction of NMR spectrogram and density for the model was achieved, and the molecular chemical formula was C199H155O36N3. The molecular structure optimization and annealing kinetics calculations are based on molecular mechanics (MM) and molecular dynamics (MD). Subsequently, a representative simplified model was constructed using the aromatic structure as the fundamental unit. On this foundation, the electrostatic potential (ESP), atomic charge distribution, and energy level orbitals were analyzed for this simplified model. The outcomes of this research can serve as an essential guide for determining the reaction order of the active categories during the low-temperature oxidation process for ZC coking coal.


Asunto(s)
Carbón Mineral , Estructura Molecular , Coque
8.
Sci Rep ; 14(1): 21591, 2024 09 16.
Artículo en Inglés | MEDLINE | ID: mdl-39284917

RESUMEN

This study introduces a novel cost-effective technique for cloning of linear DNA plasmid inserts, aiming to address the associated expenses linked with popular in vitro DNA assembly methods. Specifically, we introduce ECOLI (Efficient Cloning Of Linear Inserts), a method utilizing a PCR product-based site-directed mutagenesis. In comparison to other established in vitro DNA assembly methods, our approach is without the need for costly synthesis or specialized kits for recombination or restriction sites. ECOLI offers a fast, efficient, and economical alternative for cloning inserts up to several hundred nucleotides into plasmid constructs, thus enhancing cloning accessibility and efficiency. This method can enhance molecular biology research, as we briefly demonstrated on the Dishevelled gene from the WNT signaling pathway.


Asunto(s)
Clonación Molecular , Mutagénesis Sitio-Dirigida , Plásmidos , Plásmidos/genética , Clonación Molecular/métodos , Mutagénesis Sitio-Dirigida/métodos , Reacción en Cadena de la Polimerasa/métodos , ADN/genética
9.
J Hazard Mater ; 479: 135743, 2024 Nov 05.
Artículo en Inglés | MEDLINE | ID: mdl-39236534

RESUMEN

Direct current (DC) has promising potential for persulfate delivery and activation in heterogeneous site remediation, yet requires deeper understanding. Here, we investigated the efficiency of DC for persulfate delivery and activation and compared with alternating current (AC). While AC electric field only influenced persulfate fate by Joule heating effect, DC electric field induced electrokinetic migration of persulfate and contaminants, as well as promoted persulfate activation with Joule heating and electrochemical reactions. DC system achieved 95 % MCB removal which was 3.1 times of that in AC system using the same voltage input (60 V) with a velocity of 0.5 m/d. When the applied DC voltage increased from 20 V to 60 V (0.5-1.5 V/cm), persulfate activation pathway changed from electrode reactions to the coupled activation pathways of electrode, chemical and heat reactions, thus resulting in increasing MCB removal efficiency from 57 % (20 V) to 95 % (40 V and 60 V). The energy consumption with 40 V (11.6 kWh/g) was 2.6 times of that using 20 V (4.4 kWh/g), and dramatically increased to 11.7 times with 60 V (50.2 kWh/g). This study provides a new perspective on improving the efficiency of persulfate delivery and activation in heterogeneous sites remediation using DC-driven system.

10.
PNAS Nexus ; 3(9): pgae352, 2024 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-39228810

RESUMEN

There is an accumulation of evidence that the human gut microbiota plays a role in maintaining health, and that an altered gut microbiota (sometimes called dysbiosis) associates with risk for many noncommunicable diseases. However, the dynamics of disease-linked bacteria in the gut and other body sites remain unclear. If microbiome alterations prove causative in particular diseases, therapeutic intervention may be possible. Furthermore, microbial signature taxa have been established for the diagnosis of some diseases like colon cancer. We identified 163 disease-enriched and 98 disease-depleted gut microbiome signature taxa at species-level resolution (signature species) from 10 meta-analyses of multiple diseases such as colorectal cancer, ulcerative colitis, Crohn's disease, irritable bowel syndrome, pancreatic cancer, and COVID-19 infection. Eight signature species were enriched and nine were depleted across at least half of the diseases studied. Compared with signature species depleted in diseases, a significantly higher proportion of disease-enriched signature species were identified as extra-intestinal (primarily oral) inhabitants, had been reported in bacteremia cases from the literature, and were aerotolerant anaerobes. These findings highlight the potential involvement of oral microbes, bacteremia isolates, and aerotolerant anaerobes in disease-associated gut microbiome alterations, and they have implications for patient care and disease management.

11.
Biochem Pharmacol ; 229: 116514, 2024 Sep 03.
Artículo en Inglés | MEDLINE | ID: mdl-39236937

RESUMEN

Organic cation transporter 2 (OCT2/SLC22A2) is predominantly localized on the basolateral membranes of renal tubular epithelial cells and plays a crucial role in the renal secretion of various cationic drugs. Although variations in substrate selectivity among renal organic cation transport systems across species have been reported, the characteristics of OCT2 remain unclear. In this study, we demonstrated that atenolol, a ß1-selective adrenergic antagonist, is transported almost exclusively by human OCT2, contrasting with OCT2s from other selected species. Using chimeric constructs between human OCT2 (hOCT2) and the highly homologous monkey OCT2 (monOCT2), along with site-directed mutagenesis, we identified non-conserved amino acids Val8, Ala31, Ala34, Tyr222, Tyr245, Ala270, Ile394, and Leu503 as pivotal for hOCT2-mediated atenolol transport. Kinetic analysis revealed that atenolol was transported by hOCT2 with a 12-fold lower affinity than MPP+, a typical OCT2 substrate. The inhibitory effect of atenolol on MPP+ transport was 6200-fold lower than that observed for MPP+ on atenolol transport. Additionally, we observed weaker inhibitory effects on MPP+ transport compared to atenolol transport with ten different OCT2 substrates. Altogether, this study suggests that eight hOCT2-specific amino acids constitute the low-affinity recognition site for atenolol transport, indicating differences in OCT2-mediated drug elimination between humans and highly homologous monkeys. Our findings underscore the importance of understanding species-specific differences in drug transport mechanisms, shedding light on potential variations in drug disposition and aiding in drug development.

12.
BMC Surg ; 24(1): 252, 2024 Sep 09.
Artículo en Inglés | MEDLINE | ID: mdl-39251960

RESUMEN

BACKGROUND: Surgical site infection (SSI) is not rare after loop ileostomy reversal. This study assessed the effects of a subcutaneous closed suction drain on reducing SSIs after loop ileostomy reversal with purse-string skin closure. METHODS: This retrospective study included 229 patients who underwent loop ileostomy reversal with purse-string closure at the Pusan National University Yangsan Hospital between January 2017 and December 2021. We divided the patients into those with a subcutaneous drain (SD group) and those without it (ND group). We analyzed variables that affected SSI occurrence in both groups. RESULTS: The SD and ND groups included 109 and 120 patients, respectively. The number of incisional SSIs was significantly lower in the SD than in the ND group (0 vs. 7 events). An average of 35.7 mL of fluid was collected in the drainage bulb during hospitalization. The C-reactive protein level on postoperative day 4 was significantly lower in the SD group than in the ND group. The insertion of a subcutaneous drain was the only factor associated with a reduced incidence of SSIs (p = 0.015). CONCLUSIONS: Subcutaneous closed suction drain with purse-string skin closure in loop ileostomy reversal can reduce incisional SSI occurrence.


Asunto(s)
Ileostomía , Infección de la Herida Quirúrgica , Humanos , Estudios Retrospectivos , Infección de la Herida Quirúrgica/prevención & control , Infección de la Herida Quirúrgica/epidemiología , Infección de la Herida Quirúrgica/etiología , Masculino , Femenino , Ileostomía/métodos , Succión/métodos , Persona de Mediana Edad , Anciano , Adulto , Reoperación , Drenaje/métodos , Técnicas de Sutura
13.
BMC Surg ; 24(1): 253, 2024 Sep 10.
Artículo en Inglés | MEDLINE | ID: mdl-39256696

RESUMEN

BACKGROUND: A surgical site infection (SSI) is a postoperative infection that occurs at or near the surgical incision. SSIs significantly increase morbidity, mortality, length of hospital stay, and healthcare costs. The World Health Organization (WHO) has established hospital hygiene precaution guidelines for the prevention of SSIs, which were enhanced during the COVID-19 pandemic. The current study aims to explore the effect of the COVID-19 pandemic on SSI incidence among initially uninfected postoperative patients. We hypothesize that these enhanced precautions would reduce the incidence of SSIs. MATERIALS AND METHODS: A retrospective study comparing surgical outcomes before and during the pandemic. Patients who had abdominal surgery between June and December 2019 (Non-COVID-19) or between February and June 2020 (COVID-19) were included. The two groups were matched in a 1:1 ratio based on age, Sex, acuity (elective or emergent), surgical approach, and comorbidities. Electronic medical records were reviewed to identify SSIs and hospital readmissions within 30 days after surgery. Pearson's chi-square test and Fisher's exact test were used. RESULTS: Data was collected and analyzed from 976 patients who had surgery before the COVID-19 pandemic (non-COVID group) and 377 patients who had surgery during the pandemic (COVID group). After matching, there were 377 patients in each group. In our study, we found 23 surgical site infections (SSIs) in both laparoscopic and open surgeries. The incidence of SSIs was significantly higher in the non-COVID period compared to the COVID period [17 cases (4.5%) vs. 6 cases (1.6%), respectively, p = 0.032], especially in non-COVID open surgeries. The incidence of SSIs in laparoscopic surgeries was also higher during the non-COVID period, but not statistically significant. CONCLUSIONS: Enhanced hygiene precautions during the COVID -19 pandemic may have reduced SSIs rates following abdominal surgery.


Asunto(s)
COVID-19 , Infección de la Herida Quirúrgica , Humanos , COVID-19/prevención & control , COVID-19/epidemiología , Infección de la Herida Quirúrgica/prevención & control , Infección de la Herida Quirúrgica/epidemiología , Masculino , Femenino , Estudios Retrospectivos , Persona de Mediana Edad , Anciano , Control de Infecciones/métodos , Incidencia , Abdomen/cirugía , Pandemias , Adulto , SARS-CoV-2 , Higiene
14.
Sci Rep ; 14(1): 21998, 2024 09 23.
Artículo en Inglés | MEDLINE | ID: mdl-39313535

RESUMEN

Tiger sharks are apex predators with a circumglobal tropical and warm-temperate distribution, with a general lack of population data for the central Indian Ocean. In Fuvahmulah, Maldives, tiger sharks display frequent use of the harbour area, attracted by discarded fish waste. Here, we document the population structure, residency, and reproductive characteristics of the world's largest known tiger shark aggregation in a geographically-restricted area. Using non-invasive methods, photo identification and laser photogrammetry, we identified 239 individual tiger sharks over a 7-year study period. The aggregation was female-dominated (84.5%), with both large juveniles and adults present. Adult females were resighted over the entire study period displaying strong inter- and intra-annual site fidelity. Modelled residency using maximum likelihood methods suggests they spent 60.7 ± S.E. 7.5 days in Fuvahmulah, with a larger aggregation size, shorter residence periods and longer absence periods compared to juvenile females. Prolonged abdominal distensions of adult females indicate they likely stay near Fuvahmulah during gestation and reproduce biennially. Fuvahmulah seems to provide suitable conditions for gestation given the year-round provision of food and warm waters, exhibited by strong site fidelity and temporal residency. Our results show indications of a thriving population within the confines of protected waters.


Asunto(s)
Tiburones , Animales , Islas del Oceano Índico , Femenino , Masculino , Reproducción , Océano Índico , Maldivas
15.
J Can Chiropr Assoc ; 68(2): 160-171, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-39318847

RESUMEN

The objectives of this article, Part 2 of a two part series, are twofold: (i) To provide a narrative review of the research evidence authored by faculty of the Canadian Memorial Chiropractic College (CMCC) and; (ii) discussion of the impact this research evidence had on teaching chiropractic manual skills at CMCC and - theoretically - to the broader chiropractic educational community. Research evidence discussed are in the areas of: Experimental studies linked to biomechanics; Measuring Force - Integration of Force Sensing Table Technology (FSTT®) into technique labs; Characteristics of injuries sustained by chiropractic students during technique labs; Finding the clinical target for therapeutic intervention and; Recommendations toward a model technique curriculum. The intent of this article is for faculty at current and future accredited educational programs to incorporate this research evidence into their technique curricula and to potentially strengthen the pedagogical approach used to teach chiropractic manual skills.


L'évolution de l'enseignement des compétences manuelles en chiropratique: la deuxième partie - examen narratif et une discussion de l'impact des données probantes de recherche rédigées par le corps professoral du Canadian Memorial Chiropractic CollegeLes objectifs de cet article, la deuxième partie d'une série en deux parties, sont doubles : (i) fournir un examen narratif des données probantes de la recherche rédigées par le corps professoral du Canadian Memorial Chiropractic College (CMCC) et; (ii) discuter de l'impact de ces données probantes de la recherche sur l'enseignement des compétences manuelles en chiropratique au CMCC et - théoriquement - sur la communauté éducative en chiropratique plus large. Les preuves de recherche qui ont fait l'objet de discussions portent sur les domaines suivants : Les études expérimentales liées à la biomécanique; la mesure de la force - L'intégration de la Technologie de la table de détection de force (FSTT®) dans les laboratoires de techniques; les caractéristiques des blessures subies par les étudiants en chiropratique pendant les laboratoires de techniques; trouver la cible clinique pour l'intervention thérapeutique et; les recommandations pour un programme de technique modèle. L'objectif de cet article est que les professeurs des programmes d'enseignement accrédités actuels et futurs intègrent ces données de recherche dans leurs programmes d'études techniques et renforcent potentiellement l'approche pédagogique utilisée pour enseigner les compétences manuelles en chiropratique.

16.
Front Plant Sci ; 15: 1388040, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-39319001

RESUMEN

Pre-mRNA splicing is a fundamental process in eukaryotic gene expression, and the mechanism of intron definition, involving the recognition of the canonical GU (5'-splice site) and AG (3'-splice site) dinucleotides by splicing factors, has been postulated for most cases of splicing initiation in plants. Splice site mutations have played crucial roles in unraveling the mechanism of pre-mRNA splicing in planta. Typically, splice site mutations abolish splicing events or activate one or more cryptic splice sites surrounding the mutated region. In this report, we investigated the splicing pattern of the EGY1 gene in an Ar-ion-induced egy1-4 allele of Arabidopsis thaliana. egy1-4 has an AG-to-AC mutation in the 3'-end of intron 3, along with 4-bp substitutions and a 5-bp deletion in adjacent exon 4. RT-PCR, cDNA cloning, and amplicon sequencing analyses of EGY1 revealed that while most wild-type EGY1 mRNAs had a single splicing pattern, egy1-4 mRNAs had multiple splicing defects. Almost half of EGY1 transcripts showed 'intron retention' at intron 3, while the other half exhibited activation of 3' cryptic splice sites either upstream or downstream of the original 3'-splice site. Unexpectedly, around 8% of EGY1 transcripts in egy1-4 exhibited activation of cryptic 5'-splice sites positioned upstream of the authentic 5'-splice site of intron 3. Whole genome resequencing of egy1-4 indicated that it has no other known impactful mutations. These results may provide a rare, but real case of activation of cryptic 5'-splice sites by downstream 3'-splice site/exon mutations in planta.

17.
Sheng Wu Gong Cheng Xue Bao ; 40(9): 3072-3082, 2024 Sep 25.
Artículo en Chino | MEDLINE | ID: mdl-39319725

RESUMEN

Dextranase is an enzyme that specifically hydrolyzes the α-1, 6 glucoside bond. In order to improve the activity of dextranase from Arthrobacter oxidans KQ11, site-directed mutagenesis was used to modify the amino acids involved in the "tunnel-like binding site". A saturating mutation at position 507 was carried out on this basis. The mutant enzymes A356G, S357W, W507Y, and W507F with improved enzyme activities and catalytic efficiency were successfully obtained. Compared with wild type (WT), W507Y showed the specific activity increasing by 3.00 times, the kcat value increasing by 3.62 times, the Km value decreasing by 54%, and the catalytic efficiency (kcat/Km) increasing by 8.98 times. The three-dimensional structure analysis showed that the increase of the number of hydrogen bonds and the distance between "tunnel-like binding sites" were important factors affecting enzyme activity. Compared with WT, W507Y had a shortened distance from the residues on the other side of the "tunnel-like binding site", which made it easier to generate hydrogen binding forces. Accordingly, the substrate hydrolysis and product efflux were accelerated, which dramatically increased the enzyme activity and catalytic efficiency.


Asunto(s)
Arthrobacter , Dextranasa , Mutagénesis Sitio-Dirigida , Arthrobacter/enzimología , Arthrobacter/genética , Dextranasa/genética , Dextranasa/metabolismo , Dextranasa/química , Sitios de Unión , Mutación , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Proteínas Bacterianas/química
18.
Sheng Wu Gong Cheng Xue Bao ; 40(9): 3114-3126, 2024 Sep 25.
Artículo en Chino | MEDLINE | ID: mdl-39319728

RESUMEN

Corynebacterium glutamicum is a major workhorse in the industrial production of branched-chain amino acids (BCAAs). The acetohydroxyacid synthase (AHAS) encoded by ilvBN is a key enzyme in the biosynthesis of BCAAs. Enhancing AHAS expression is essential for engineering BCAA producers. However, at present, the available studies only used limited promoters to regulate AHAS expression, which is insufficient for achieving efficient regulation. Herein, we first employed a previously developed reporter system to screen out a strong constitutive promoter PgpmA* from six candidate promoters for expressing ilvBN. PgpmA* showcased the expression strength 23.3-fold that of the native promoter PilvBN. Moreover, three synthetic RBS libraries based on the promoter PgpmA* were constructed and evaluated by plate fluorescence imaging. The results revealed that "R(9)N(6)" was the best mutant library. A total of 36 RBS mutants with enhanced strength were further screened by evaluation in 96-deep-well plates, and the highest strength reached up to 62.3-fold that of PilvBN. Finally, the promoter PgpmA* was combined with three RBS mutants (WT, RBS18, and RBS36) to fine-tune the expression of ilvBNS155F for L-valine biosynthesis, respectively. Increased expression strength led to enhanced L-valine production, with titers of 1.17, 1.38, and 2.29 g/L, respectively. The combination of RBS18 strain with the further overexpression of ilvC produced 7.57 g/L L-valine. The regulatory elements obtained in this study can be utilized to modulate AHAS expression for BCAA production in C. glutamicum. Additionally, this strategy can guide the efficient expression regulation of other key enzymes.


Asunto(s)
Acetolactato Sintasa , Aminoácidos de Cadena Ramificada , Corynebacterium glutamicum , Regulación Bacteriana de la Expresión Génica , Regiones Promotoras Genéticas , Corynebacterium glutamicum/genética , Corynebacterium glutamicum/metabolismo , Aminoácidos de Cadena Ramificada/biosíntesis , Aminoácidos de Cadena Ramificada/metabolismo , Aminoácidos de Cadena Ramificada/genética , Acetolactato Sintasa/genética , Acetolactato Sintasa/metabolismo , Ingeniería Metabólica/métodos , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo
19.
Emerg Infect Dis ; 30(11)2024 Sep 25.
Artículo en Inglés | MEDLINE | ID: mdl-39322417

RESUMEN

We traced back a nationwide outbreak of human listeriosis in Switzerland to a persisting production line contamination of a factory producing baker's yeast with Listeria monocytogenes serotype 1/2a sequence type 3141. We used whole-genome sequencing to match clinical isolates to isolates from product samples.

20.
Artículo en Inglés | MEDLINE | ID: mdl-39322461

RESUMEN

Gestational trophoblastic disease (GTD), comprising hydatidiform moles (HM) and gestational trophoblastic tumors (GTT), is extremely rare. HM originate from villous trophoblast and are considered preneoplastic. GTT originate from the intermediate, largely extravillous trophoblast and includes choriocarcinoma, placental site trophoblastic tumor, epitheloid trophoblastic tumor, and mixed trophoblastic tumor. The abnormal (non-molar) villous lesions, non-malignant tumour-like conditions, and non-gestational tumors add to the diagnostic dilemma. The correct diagnosis and classification of these rare conditions are important. This review intends to provide an update on changes in the World Health Organization classification and focusses on the morphologic aspects in diagnosis of GTD.

SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA