RESUMO
Spinal muscular atrophy (SMA) is a neuromuscular disorder with an autosomal recessive inheritance pattern. Patients with severe symptoms may suffer respiratory failure, leading to death. The homozygous deletion of exon 7 in the SMN1 gene accounts for nearly 95% of all cases. Population carrier screening for SMA and prenatal diagnosis by amniocentesis for high-risk couples can assist in identifying the risk of fetal disease. We provided the SMA carrier screening process to 55,447 pregnant women in Yancheng from October 2020 to December 2022. Among them, 8185 participated in this process, with a participation rate of around 14.76% (95% CI 14.47-15.06%). Quantitative real-time polymerase chain reaction (qPCR) was used to detect deletions of SMN1 exons 7 and 8 (E7, E8) in screened pregnant women. 127 were identified as carriers (111 cases of E7 and E8 heterozygous deletions, 15 cases of E7 heterozygous deletions, and 1 case of E7 heterozygous deletions and E8 homozygous deletions), resulting in a carrying rate of around 1.55% (95% CI 1.30-1.84%). After genetic counseling, 114 spouses of pregnant women who tested positive underwent SMA carrier screening; three of them were screened as SMA carriers. Multiplexed ligation-dependent probe amplification (MLPA) was used for the prenatal diagnosis of the fetuses of high-risk couples. Two of them exhibited two copies of SMN1 exon 7 (normal), and the pregnancy was continued; one exhibited no copies of SMN1 exon 7 and exon 8 (SMA patient), and the pregnancy was terminated. Analyzing SMN1 mutations in Yancheng and provide clinical evidence for SMA genetic counseling and birth defect prevention. Interventional prenatal diagnosis for high-risk families can promote informed reproductive selection and prepare for the fetus's early treatment.
RESUMO
Collagen type I alpha1 (COL1A1) has been found to be abnormal expressed in oral squamous cell carcinoma (OSCC) tissues, but its role and mechanism in OSCC need to be further elucidated. The expression levels of COL1A1 and methyltransferase-like 3 (METTL3) were measured by quantitative real-time PCR and western blot. Cell growth and metastasis were determined by CCK8, colony formation, EdU, flow cytometry and transwell assays. MeRIP, Co-IP and dual-luciferase reporter assays were performed to explore the interplay of COL1A1 and METTL3. COL1A1 mRNA stability was confirmed by Actinomycin D assay. Mice xenograft models were constructed to perform in vivo experiments. COL1A1 and METTL3 were upregulated in OSCC. COL1A1 knockdown suppressed OSCC cell growth and metastasis, while its overexpression had an opposite effect. The stability of COL1A1 mRNA was regulated by the m6A methylation of METTL3. METTL3 overexpression promoted OSCC cell growth and metastasis, and its knockdown-mediated OSCC cell function inhibition could be abolished by COL1A1 overexpression. Besides, silencing of METTL3 reduced OSCC tumor growth by reducing COL1A1 expression. METTL3-stabilized COL1A1 promoted OSCC progression, providing an exact molecular target for the treatment of OSCC.
RESUMO
Zn metal as a promising anode of aqueous batteries faces severe challenges from dendrite growth and side reactions. Here, tetraphenylporphyrin tetrasulfonic acid (TPPS) is explored as an electrolyte additive for advanced Zn anodes. It is interesting to note that TPPS spontaneously assembles into unique aggregates. As they adsorb on the Zn anode, the aggregates enhance the resistance to electrolyte percolation and dendrite growth compared to single molecules. Meanwhile, TPPS facilitates anion association in the solvation sheath of Zn2+, and boosts the transference number of Zn2+ up to 0.95. Therefore, anion-related side reactions and anion-induced electrode overpotentials are reduced accordingly. In this context, the electrolyte containing TPPS exhibits excellent electrochemical performance. Even under a high loading of MnO2 (25â mg cm-2), a limited Zn supply (N/P ratio=1.7), and a lean electrolyte (15â µL mAh-1), the full cells still represent a higher cumulative capacity compared to the reported data. The advantages of this electrolyte are also adapted to other cathode materials. The pouch cells of Zn||NaV3O8 â 1.5H2O realize a capacity of ~0.35â Ah at 0.4â C under harsh conditions.
RESUMO
Five undescribed steroids and one sesquiterpene, named Aglaians A-F, along with sixteen known analogs, have been isolated from the branches and leaves of Aglaia lawii. Its structure was elucidated by IR, HRESIMS, 1D and 2D NMR, quantum-chemical calculations, electronic circular dichroism (ECD) calculations, and single-crystal X-ray diffraction analysis. The cytotoxic and antibacterial activities of six human tumor cell lines were evaluated (MDA-MB-231, MCF-7, Ln-cap, A549, HeLa, and HepG-2), and four strains of bacteria (Bacterium subtilis, Phytophthora cinnamomic, Acrogenic bacterium, and Ralstonia solanacearum). The bioassay results indicated that compounds 3 and 5 exhibited moderate antitumor activity with IC50 values ranging from 16.72 to 36.14 µM. Furthermore, compounds 3-5 possess antibacterial activities against four bacteria with MIC values of 25-100 µM.
Assuntos
Aglaia , Sesquiterpenos , Humanos , Esteroides/farmacologia , Antibacterianos/farmacologia , Linhagem Celular Tumoral , Sesquiterpenos/farmacologiaRESUMO
Limosilactobacillus reuteri 121 4,6-α-glucanotransferase GtfB (Lr 121 GtfB), belonging to glycoside hydrolase family 70 (GH70), synthesizes linear isomalto/malto polysaccharides having (α1â6) linkages attached to the nonreducing ends of (α1â4) linked maltose oligosaccharide segments using starch or maltodextrin as a substrate. Since Lr 121 GtfB has low catalytic activity and efficiency, it leads to substrate regeneration and reduced substrate utilization. In this study, we superimposed the crystal structure of Lr 121 GtfB-ΔNΔV with that of L. reuteri NCC 2613 GtfB-ΔNΔV (Lr 2613 GtfB-ΔNΔV) to identify the acceptor binding subsites +1 to +3 and constructed five single-residue mutants and a random mutagenesis of N1019. Compared with the wild-type, N1019D Lr 121 GtfB-ΔN did not alter the product specificity, increased the catalytic activity and efficiency by 420 and 590%, respectively, and maintained >80% relative activity in the pH 3.5-6.5 interval. The findings will contribute to the industrial application of Lr 121 GtfB and provide new solutions for starch synthesis of higher value derivatives.
Assuntos
Sistema da Enzima Desramificadora do Glicogênio , Limosilactobacillus reuteri , Sistema da Enzima Desramificadora do Glicogênio/química , Amido/metabolismo , OligossacarídeosRESUMO
CsPbBr3 has received more and more attention in the field of optoelectronic devices due to its excellent stability. To address the cost and environmental concerns associated with the use of toxic methanol, water has been explored as a substitute solvent for CsBr in the preparation of CsPbBr3 perovskite solar cells (PSCs). In this study, we utilized methanol as an anti-solvent of the CsBr/H2O solution to regulate the detrimental effects of water on the CsPbBr3 film and control the crystallization process. From results of the experiment, it was found that methanol anti-solvent treatment greatly improved the crystallization of the CsPbBr3 film, increased the grain size, and reduced the defect density. After the introduction of methanol anti-solvent treatment, the power conversion efficiency (PCE) increased from 6.09% to 7.91%, while the open-circuit voltage (Voc) increased from 1.18 V to 1.39 V. Furthermore, we incorporated 2-hydroxyethylurea into the CsPbBr3 PSCs to improve the wettability of PbBr2 towards the CsBr/H2O solution and ensure the formation of pure-phase CsPbBr3 films. The introduction of 2-hydroxyethylurea resulted in an additional increase in Voc from 1.19 V to 1.42 V. The PCE further improved from 6.56% to 8.62% after methanol anti-solvent treatment. These results demonstrate that methanol treatment effectively addresses the low Voc issue observed in CsPbBr3 PSCs prepared with water as a solvent. Importantly, this approach significantly reduces the reliance on methanol compared to conventional fabrication methods for CsPbBr3 PSCs. Overall, this work presents a promising pathway for achieving high Voc and efficiency in CsPbBr3 PSCs by utilizing water as a solvent.
RESUMO
Due to its high efficacy as a wide-spectrum disinfectant and its potential for the degradation of pollutants and pesticides, ozone has broad application prospects in agricultural production. In this study, micro/nano bubble technology was applied to achieve a saturation state of bubble nutrient solution, including micro-nano oxygen (O2 group) and micro-nano ozone (O3 group) bubble nutrient solutions. The effects of these solutions on lettuce physiological indices as well as changes in the microbial community within the rhizosphere substrate were studied. The application of micro/nano (O2 and O3) bubble nutrient solutions to substrate-cultured lettuce plants increased the amount of dissolved oxygen in the nutrient solution, increased the lettuce yield, and elevated the net photosynthetic rate, conductance of H2O and intercellular carbon dioxide concentration of lettuce plants. Diversity analysis of the rhizosphere microbial community revealed that both the abundance and diversity of bacterial and fungal communities in the substrate increased after plant cultivation and decreased following treatment with micro/nanobubble nutrient solutions. RDA results showed that the microbial community in the S group was positively associated with EC, that in the CK and O2 groups exhibited a positive correlation with SC, and that in the O3 group displayed a positive correlation with CAT and POD. Overall, the implementation of micro/nanobubble generation technology in soilless substrates can effectively increase the lettuce growth and yield, and O3 had a more pronounced effect on lettuce yield and quality and the microbial community structure in the substrate than O2. Our study would provide a reference and theoretical basis for developing sustainable and green technology for promoting lettuce production and can be a promising alternative to conventional methods for improving crop yields.
RESUMO
The large thixotropy of the starch-thickened foods is often unfavorable in many applications. This study examined the contribution of the proportion of amylopectin chain length to time-dependence of starch gels. The α-amylase (AM) from Bacillus stearothermophilus and maltogenic α-amylase (MA) from Bacillus subtilis were used to trim amylopectin in different reaction patterns. HPLC, HPAEC and IBC data suggested AM attacked B-chains (DP 12-36), causing an increment in number of the chains with DP 6-12, whereas MA primarily trimmed the short B-chains (DP 12-18) and partial A-chains (DP 9-12) to generate short chains with DP 6-9. Interestingly, the recovery of AM-gels was faster than MA-gels at the same degree of hydrolysis when subjected to shear according to the linear correlation analysis. When releasing the same mass of sugar, shortening of the long internal chains played an important role in reducing time dependence of starch gel rather than the external side chains. Possible models were proposed to illustrate the differences in the mechanism of rapid-recovery caused by different side-chain distributions. The outcome provided a new perspective to regulate the thixotropy behavior of starch through enzyme strategies in the granular state.
Assuntos
Amilopectina , Amido , Zea mays , alfa-Amilases , alfa-Amilases/metabolismo , alfa-Amilases/química , Zea mays/química , Amido/química , Amido/metabolismo , Amilopectina/química , Hidrólise , Géis/química , Geobacillus stearothermophilus/enzimologia , Bacillus subtilis/enzimologiaRESUMO
INTRODUCTION: The present research focused on the function of lncRNA taurine upregulated 1 (TUG1) in a rat neuropathic pain (NP) model constructed by chronic contractile injury (CCI). MATERIAL AND METHODS: Construction of the NP rat model was performed by CCI surgery. Paw withdrawal threshold (PWT) and paw withdrawal latency (PWL) were applied to examine the NP behavior. RT-qPCR was established to explore the levels of TUG1, microRNA (miR)-29b-3p, and HMGB1. ELISA was carried out to evaluate the concentrations of interleukin (IL)-6, IL-1ß, tumor necrosis factor α (TNF-α), IL-4, and IL-6. The underlying mechanisms of TUG1 were explored by RNA-binding protein immunoprecipitation (RIP) and dual-luciferase reporter (DLR) assay. RESULTS: TUG1 and HMGB1 were statistically elevated in the tissue of CCI rats, while miR-29b-3p was reduced. TUG1 competitively binds to miR-29b-3p to upregulate HMGB1 levels. Suppression of TUG1 persistently decreased PWL and PWT along with increased frequency of paw-lifting, whereas this alleviation was typically rescued by the abrogated miR-29b-3p. Analogously, knockdown of TUG1 inhibited CCI-induced overproduction of IL-6, IL-1ß, and TNF-α, and reduction of IL-4 and IL-6, but this inhibition was partially abrogated by the reduction of miR-29b-3p. CONCLUSIONS: Suppression TUG1 can alleviate NP hypersensitivity and neuroinflammation in CCI rats by competitively binding miR-29b-3p to weaken HMGB1.
RESUMO
Starch-converting α-glucanotransferases are efficient enzymatic toolkits for the biosynthesis of diverse α-glucans, which hold vast application potential in the food industry. In this work, we identified a novel GtfB protein from Fructilactobacillus sanfranciscensis TMW11304 (FsTMW11304 GtfB) in NCBI. Although this enzyme was highly conserved in motifs I-IV with those isomalto-maltopolysaccharides (IMMPs)-producing GtfB α-glucanotransferases, it possessed distinct deletions and mutations in two crucial loops shaping the active site. Hence, unlike those GtfB enzymes, FsTMW11304 GtfB not only exhibited excellent 4,6-α-glucanotransferase activity on amylose to generate atypically low-molecular-weight IMMPs with consecutive linear (α1 â 6) linkages up to 48 %, but also held good capability towards branched substrates. Besides, compared with the control, the treatment by FsTMW11304 GtfB reduced the storage/loss modulus of granular and gelatinized tapioca starches (TS) by 12.0 %/17.9 % and 91.4 %/82.9 %, respectively, indicating that the rigidity of the gel structure was attenuated to different degrees in the two reaction systems. Furthermore, the setback viscosity observed in the gelatinized TS modified by FsTMW11304 GtfB was only 5 % of that observed in the control group, suggesting the short-term anti-retrogradation property has been substantially improved. Thus, FsTMW11304 GtfB represents a meaningful addition to the α-glucanotransferases in GH70 family, which expands the repertoire of diverse α-glucans synthesized from starch and facilitates the understanding of the structure-function relationship of the GtfB α-glucanotransferases.
Assuntos
Lactobacillus , Manihot , Amido , Amido/metabolismo , Manihot/metabolismo , Viscosidade , Glucanos/química , AmiloseRESUMO
Starch-converting α-glucanotransferases of glycoside hydrolase family 70 (GH70) are promising enzymatic tools for the production of diverse α-glucans with (potential) commercial applications in food and health and as biomaterials. In this study, a novel GtfB enzyme from Weissella confusa MBF8-1 was screened in the National Center for Biotechnology Information (NCBI) nonredundant protein database. The enzyme (named WcMBF8-1 GtfB) displayed high conservation in motifs I-IV with other GtfB enzymes but possessed unique variations in several substrate-binding residues. Structural characterizations of its α-glucan products revealed that WcMBF8-1 GtfB exhibited an atypical 4,6-α-glucanotransferase activity and was capable of catalyzing, by cleaving off (α1 â 4)-linkages in starch-like substrates and the synthesis of linear (α1 â 6) linkages and (α1 â 4,6) branching points. The product specificity enlarges the diversity of α-glucans and facilitates recognition of the determinants of the linkage specificity in GtfB enzymes. Furthermore, the contents of slowly digestible starch and resistant starch of granular corn starches, modified by WcMBF8-1 GtfB, increased by 6.7%, which suggested the potential value for the utilization of WcMBF8-1 GtfB to prepare "clean-label" starch ingredients with improved functional attributes.
Assuntos
Amilose , Sistema da Enzima Desramificadora do Glicogênio , Zea mays , Amilose/metabolismo , Zea mays/metabolismo , Amido/metabolismo , Glucanos/químicaRESUMO
Zn anodes of aqueous batteries face severe challenges from side reactions and dendrite growth. Here, triethanolamine (TEOA) is developed as an electrolyte additive to address these challenges. It enhances the exposure of Zn(002) and diminishes the change in pH. Therefore, the electrolyte containing TEOA shows improved electrochemical performance.
RESUMO
The recently characterized Limosilactobacillus reuteri N1 GtfB (LrN1 GtfB) from glycoside hydrolase family 70 is a novel 4,6-α-glucanotransferase acting on starch/maltooligosaccharides with high enzyme activity and soluble protein yield (in heterogenous system). In this study, the influence of the treatment by LrN1 GtfB on the fine structure and functional characteristics of three maize starches were furtherly investigated and elucidated. Due to the treatment of LrN1 GtfB, the starch molecules were transformed into reuterans containing linear and branched (α1 â 6) linkages with notably smaller molecular weight and shorter chain length. Moreover, the (α1 â 6) linkage ratios in the GtfB-modified high-amylose maize starch (GHMS)/normal maize starch (GNMS)/waxy maize starch (GWMS) increased by 18.3 %/12.6 %/9.0 % as compared to their corresponding controls. In vitro digestibility experiment revealed that the resistant starch content of GHMS, GNMS and GWMS increased by 16 %, 18 % and 25 % as compared to the starch substrates. Furthermore, the butyric acid yielded from GHMS, GNMS and GWMS in the in vitro fermentation experiments were 1.4, 1.5 and 1.4 times higher than those of commercial galactose oligosaccharides. These results indicated that the highly-branched short-clustered reuteran synthesized by LrN1 GtfB might serve as novel potential prebiotics, and provide insights for the synthesis of promising prebiotic dietary fiber from starch.
Assuntos
Limosilactobacillus reuteri , Prebióticos , Amido , Zea mays , Zea mays/química , Amido/química , Amido/metabolismo , Oligossacarídeos/químicaRESUMO
The accumulation of polyurethane plastics (PU-PS) in the environment is on the rise, posing potential risks to the health and function of ecosystems. However, little is known about the degradation behavior of PU-PS in the environment, especially water environment. To address this knowledge gap, we investigated and isolated a degrading strain of Streptomyces sp. B2 from the surface of polyurethane coatings. Subsequently, a photoreactor was employed to simulate the degradation process of bio-based polyurethane (BPU) and petroleum-based polyurethane (PPU) under three conditions, including single microorganism (SM), single light exposure (SL), and combined light exposure/microorganism action (ML) in aqueous solution. The results indicated that PU-PS mainly relies on biodegradation, with the highest degradation rate observed after 28 d under SM condition (BPU 5.69 %; PPU 5.25 %). SL inhibited microbial growth and degradation, with the least impact on plastic degradation. Microorganisms colonized the plastic surface, secreting relevant hydrolytic enzymes and organic acids into the culture medium, providing a negative charge. The carbon chains were broken and aged through hydrogen peroxide induction or attack by oxygen free radicals. This process promoted the formation of oxidized functional groups such as OH and CO, disrupting the polymer's structure. Consequently, localized fragmentation and erosion of the microstructure occurred, resulting in the generation of secondary microplastic (MPs) particles, weight loss of the original plastic, increased surface roughness, and enhanced hydrophilicity. Additionally, BPU exhibited greater degradability than PPU, as microorganisms could utilize the produced fatty acids, which promoted their reproduction. In contrast, PPU degradation generated a large amount of isocyanate, potentially toxic to cells and inhibiting biodegradation. This study unveils the significant role of microorganisms in plastic degradation and the underlying degradation mechanisms of BPU, providing a novel strategy for polyurethane degradation and valuable information for comprehensive assessment of the behavior and fate of MPs in the environment.
Assuntos
Biodegradação Ambiental , Luz , Poliuretanos , Poliuretanos/química , Plásticos , Streptomyces/metabolismoRESUMO
The function of polysaccharides is intimately associated with their size, which is largely determined by the processivity of transferases responsible for their synthesis. A tunnel active center architecture has been recognized as a key factor that governs processivity of several glycoside hydrolases (GHs), e.g., cellulases and chitinases. Similar tunnel architecture is also observed in the Limosilactobacillus reuteri 121 GtfB (Lr121 GtfB) α-glucanotransferase from the GH70 family. The molecular element underpinning processivity of these transglucosylases remains underexplored. Here, we report the synthesis of the smallest (α1 â 4)-α-glucan interspersed with linear and branched (α1 â 6) linkages by a novel 4,6-α-glucanotransferase from L. reuteri N1 (LrN1 GtfB) with an open-clefted active center instead of the tunnel structure. Notably, the loop swapping engineering of LrN1 GtfB and Lr121 GtfB based on their crystal structures clarified the impact of the loop-mediated tunnel/cleft structure at the donor subsites -2 to -3 on processivity of these α-glucanotransferases, enabling the tailoring of both product sizes and substrate preferences. This study provides unprecedented insights into the processivity determinants and evolutionary diversification of GH70 α-glucanotransferases and offers a simple route for engineering starch-converting α-glucanotransferases to generate diverse α-glucans for different biotechnological applications.
Assuntos
Proteínas de Bactérias , Glucanos , Limosilactobacillus reuteri , Glucanos/química , Glucanos/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Limosilactobacillus reuteri/enzimologia , Limosilactobacillus reuteri/genética , Limosilactobacillus reuteri/química , Domínio Catalítico , Glucosiltransferases/química , Glucosiltransferases/genética , Glucosiltransferases/metabolismo , Engenharia de Proteínas , Sistema da Enzima Desramificadora do Glicogênio/genética , Sistema da Enzima Desramificadora do Glicogênio/metabolismo , Sistema da Enzima Desramificadora do Glicogênio/químicaRESUMO
Autoimmune related kidney diseases (ARKDs), including minimal change nephropathy (MCN), membranous nephropathy (MN), IgA nephropathy (IgAN), and lupus nephritis (LN), significantly affect renal function. These diseases are characterized by the formation of local immune complexes and the subsequent activation of the complement system, leading to kidney damage and proteinuria. Despite the known patterns of glomerular injury, the specific molecular mechanisms that contribute to renal tubular damage across ARKDs remain underexplored. Laser capture microdissection and liquid chromatography-tandem mass spectrometry (LC-MS/MS) were used to conduct a comparative proteomic analysis of renal tubular tissues from formalin-fixed paraffin-embedded samples. The cohort comprised of 10 normal controls (NC), 5 MCN, 4 MN, 17 IgAN, and 21 LN patients. Clinical parameters and histopathological assessments were integrated with proteomic findings to comprehensively investigate underlying pathogenic processes. Clinical evaluation indicated significant glomerular damage, as reflected by elevated urinary protein levels and reduced plasma albumin levels in patients with ARKD. Histological analyses confirmed varying degrees of tubular damage and deposition of immune complexes. Proteomic analyses identified significant changes in protein expression, particularly in complement components (C3, C4A, C4B, C8G, CFB, and SERPINA1) and mitochondrial proteins (ATP5F1E and ATP5PD), highlighting the common alterations in the complement system and mitochondrial proteins across ARKDs. These alterations suggest a novel complement-mitochondrial-epithelial-mesenchymal transition (EMT) pathway axis that contributes to tubular damage in ARKDs. Notably, significant alterations in CFB in tubular ARKD patients were revealed, implicating it as a therapeutic target. This study underscores the importance of complement activation and mitochondrial dysfunction in the pathogenesis of ARKDs, and proposes CFB as a potential therapeutic target to inhibit complement activation and mitigate tubular damage. Future research should validate the complement-mitochondrial-EMT pathway axis and explore the effects and mechanisms of CFB inhibitors in alleviating ARKD progression.
Assuntos
Ativação do Complemento , Mitocôndrias , Proteômica , Humanos , Proteômica/métodos , Feminino , Masculino , Adulto , Mitocôndrias/metabolismo , Pessoa de Meia-Idade , Nefropatias/metabolismo , Nefropatias/patologia , Nefropatias/imunologia , Doenças Autoimunes/metabolismo , Doenças Autoimunes/imunologia , Doenças Autoimunes/patologia , Túbulos Renais/metabolismo , Túbulos Renais/patologia , Espectrometria de Massas em Tandem , Microdissecção e Captura a Laser , Proteínas do Sistema Complemento/metabolismo , Cromatografia LíquidaRESUMO
Diabetic nephropathy (DN) remains the primary cause of end-stage renal disease (ESRD), warranting equal attention and separate analysis of glomerular, tubular, and interstitial lesions in its diagnosis and intervention. This study aims to identify the specific proteomics characteristics of DN, and assess changes in the biological processes associated with DN. 5 patients with DN and 5 healthy kidney transplant donor control individuals were selected for analysis. The proteomic characteristics of glomeruli, renal tubules, and renal interstitial tissue obtained through laser capture microscopy (LCM) were studied using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Significantly, the expression of multiple heat shock proteins (HSPs), tubulins, and heterogeneous nuclear ribonucleoproteins (hnRNPs) in glomeruli and tubules was significantly reduced. Differentially expressed proteins (DEPs) in the glomerulus showed significant enrichment in pathways related to cell junctions and cell movement, including the regulation of actin cytoskeleton and tight junction. DEPs in renal tubules were significantly enriched in glucose metabolism-related pathways, such as glucose metabolism, glycolysis/gluconeogenesis, and the citric acid cycle. Moreover, the glycolysis/gluconeogenesis pathway was a co-enrichment pathway in both DN glomeruli and tubules. Notably, ACTB emerged as the most crucial protein in the protein-protein interaction (PPI) analysis of DEPs in both glomeruli and renal tubules. In this study, we delve into the unique proteomic characteristics of each sub-region of renal tissue. This enhances our understanding of the potential pathophysiological changes in DN, particularly the potential involvement of glycolysis metabolic disorder, glomerular cytoskeleton and cell junctions. These insights are crucial for further research into the identification of disease biomarkers and the pathogenesis of DN.
Assuntos
Nefropatias Diabéticas , Rim , Microdissecção e Captura a Laser , Proteoma , Proteômica , Espectrometria de Massas em Tandem , Humanos , Nefropatias Diabéticas/metabolismo , Nefropatias Diabéticas/patologia , Proteômica/métodos , Microdissecção e Captura a Laser/métodos , Masculino , Pessoa de Meia-Idade , Feminino , Espectrometria de Massas em Tandem/métodos , Rim/química , Rim/metabolismo , Rim/patologia , Proteoma/análise , Proteoma/metabolismo , Cromatografia Líquida de Alta Pressão/métodos , Adulto , IdosoRESUMO
Prunus conradinae (subgenus Cerasus, Rosaceae) is a significant germplasm resource of wild cherry blossom in China. To ensure the comprehensiveness of this study, we used a large sample size (12 populations comprising 244 individuals) which involved the fresh leaves of P. conradinae in Eastern, Central, and Southwestern China. We combined morphological and molecular evidence (three chloroplast DNA (cpDNA) sequences and one nuclear DNA (nr DNA) sequence) to examine the population of P. conradinae variation and differentiation. Our results revealed that Central, East, and Southwest China are important regions for the conservation of P. conradinae to ensure adequate germplasm resources in the future. We also found support for a new variant, P. conradinae var. rubrum. We observed high genetic diversity within P. conradinae (haplotype diversity [Hd] = 0.830; ribotype diversity [Rd] = 0.798), with novel genetic variation and a distinct genealogical structure among populations. There was genetic variation among populations and phylogeographic structure among populations and three geographical groups (Central, East, and Southwest China). The genetic differentiation coefficient was the lowest in the Southwest region and the gene exchange was obvious, while the differentiation was obvious in Central China. In the three geographic groups, we identified two distinct lineages: an East China lineage (Central China and East China) and a Southwest China lineage ((Central China and Southwest China) and East China). These two lineages originated approximately 4.38 million years ago (Mya) in the early Pliocene due to geographic isolation. P. conradinae expanded from Central China to East China at 3.32 Mya (95% HPD: 1.12-5.17 Mya) in the Pliocene. The population of P. conradinae spread from East China to Southwest China, and the differentiation time was 2.17 Mya (95% (HPD: 0.47-4.54 Mya), suggesting that the population of P. conradinae differentiated first in Central and East China. The population of P. conradinae experienced differentiation from Central China to Southwest China around 1.10 Mya (95% HPD: 0.11-2.85 Mya) during the early Pleistocene of the Quaternary period. The southeastern region of East China, near Mount Wuyi, likely serves as a refuge for P. conradinae. This study establishes a theoretical foundation for the classification, identification, conservation, and exploitation of germplasm resources of P. conradinae.
RESUMO
Prunustongmuensis, a new species of cherry blossom, is described and illustrated from Wuyishan National Park, southeast China. This species is characterized by its tubular to nearly bottle-shaped receptacles and dark purple drupes. It can be distinguished from other wild cherry trees by its flowers and leaves, reddish brown young leaves, presence of 1-2 glands at the base of leaves, petioles densely covered with yellowish brown villi, longer pedicels (0.6-2.5 cm), villous pistil, and dark purple drupes. In the present study, we conducted a comprehensive morphological study based on specimens of the new species and its morphologically close species, field observations, and examination of pollen morphology. In addition, our phylogenetic analysis based on the complete plastid genome sequences further confirms the status of the new species and indicates that it is closely related to Prunusclarofolia, however, it notably differs in leaf shape, size, petiole villus color, gland location, timing of flower and leaf openings, and reflexed or spread sepals, as well as drupe color.
RESUMO
α-Glucanotransferases of the CAZy family GH70 convert starch-derived donors to industrially important α-glucans. Here, we describe characteristics of a novel GtfB-type 4,6-α-glucanotransferase of high enzyme activity (60.8 U mg-1) from Limosilactobacillus reuteri N1 (LrN1 GtfB), which produces surprisingly large quantities of soluble protein in heterologous expression (173 mg pure protein per L of culture) and synthesizes the reuteran-like α-glucan with (α1 â 6) linkages in linear chains and branch points. Protein structural analysis of LrN1 GtfB revealed the potential crucial residues at subsites -2â¼+2, particularly H265, Y214, and R302, in the active center as well as previously unidentified surface binding sites. Furthermore, molecular dynamic simulations have provided unprecedented insights into linkage specificity hallmarks of the enzyme. Therefore, LrN1 GtfB represents a potent enzymatic tool for starch conversion, and this study promotes our knowledge on the structure-function relationship of GH70 GtfB α-glucanotransferases, which might facilitate the production of tailored α-glucans by enzyme engineering in future.