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Plant Biotechnology Journal Oct 2023High-molecular-weight glutenin subunits (HMW-GS), a major component of seed storage proteins (SSP) in wheat, largely determine processing quality. HMW-GS encoded by...
High-molecular-weight glutenin subunits (HMW-GS), a major component of seed storage proteins (SSP) in wheat, largely determine processing quality. HMW-GS encoded by GLU-1 loci are mainly controlled at the transcriptional level by interactions between cis-elements and transcription factors (TFs). We previously identified a conserved cis-regulatory module CCRM1-1 as the most essential cis-element for Glu-1 endosperm-specific high expression. However, the TFs targeting CCRM1-1 remained unknown. Here, we built the first DNA pull-down plus liquid chromatography-mass spectrometry platform in wheat and identified 31 TFs interacting with CCRM1-1. TaB3-2A1 as proof of concept was confirmed to bind to CCRM1-1 by yeast one hybrid and electrophoretic mobility shift assays. Transactivation experiments demonstrated that TaB3-2A1 repressed CCRM1-1-driven transcription activity. TaB3-2A1 overexpression significantly reduced HMW-GS and other SSP, but enhanced starch content. Transcriptome analyses confirmed that enhanced expression of TaB3-2A1 down-regulated SSP genes and up-regulated starch synthesis-related genes, such as TaAGPL3, TaAGPS2, TaGBSSI, TaSUS1 and TaSUS5, suggesting that it is an integrator modulating the balance of carbon and nitrogen metabolism. TaB3-2A1 also had significant effects on agronomic traits, including heading date, plant height and grain weight. We identified two major haplotypes of TaB3-2A1 and found that TaB3-2A1-Hap1 conferred lower seed protein content, but higher starch content, plant height and grain weight than TaB3-2A1-Hap2 and was subjected to positive selection in a panel of elite wheat cultivars. These findings provide a high-efficiency tool to detect TFs binding to targeted promoters, considerable gene resources for dissecting regulatory mechanisms underlying Glu-1 expression, and a useful gene for wheat improvement.
Topics: Triticum; Proteome; Transcription Factors; Glutens; Promoter Regions, Genetic; Edible Grain; Starch; Molecular Weight
PubMed: 37381172
DOI: 10.1111/pbi.14103 -
BMC Musculoskeletal Disorders Feb 2024Knee osteoarthritis is severe progressive and most commonly diagnosed articular disease and its incidence is increasing around the world depending on age. This...
BACKGROUND
Knee osteoarthritis is severe progressive and most commonly diagnosed articular disease and its incidence is increasing around the world depending on age. This pathologic condition which limits daily activity of patients can be characterized by degeneration of cartilage and inflammation. Although non-steroidal anti-inflammatory (NSAII) agents and other analgesics are routinely used treatment options, the potential effects of intraarticular injections including hyaluronic acid (HA) have also been demonstrated by various studies. However, few studies compare the efficacy of a single high molecular weight (HMW) high dose and a triple HMW low dose. This study aimed to compare the efficacy of single high molecular weight (HMW) high dose (2 mL / 60 mg) and triple HMW low dose (2 mL /30 mg) intra-articular injection of HA in knee osteoarthritis (OA) patients by evaluating function and pain parameters during 12 months.
METHODS
This is a single-center, retrospective clinical study that included and involved 128 patients. Group I (n=64) patients received triple 30 mg HA injections (SEMICAL®) with one-week intervals, while Group II (n=64) patients received a single 60 mg HA injection (SEMICAL®). Lequesne Index, WOMAC and VAS scores were recorded to assess pain and function during a 12-month period.
RESULTS
There was no significant difference in characteristics of patient demographics. Our finding indicate that WOMAC, VAS score, and Lequesne Index values during follow-up visits exhibited a decrease, signifying improvement in the clinical condition. Notably, scores were significantly more favorable with the 30 mg of HA injection compared to the 60 mg of HA injection.
CONCLUSION
This study suggests that the triple low-dose injection of HMW HA is more effective in improving WOMAC, VAS scores and Lequesne Index values than a single high-dose injection.
Topics: Humans; Hyaluronic Acid; Osteoarthritis, Knee; Viscosupplements; Molecular Weight; Retrospective Studies; Treatment Outcome; Pain; Injections, Intra-Articular; Anti-Inflammatory Agents, Non-Steroidal
PubMed: 38383379
DOI: 10.1186/s12891-024-07200-y -
Obstetric Medicine Dec 2023In pregnant women, low molecular weight heparin is recommended as the preferred agent for venous thromboembolism prophylaxis and treatment. Despite their widespread...
In pregnant women, low molecular weight heparin is recommended as the preferred agent for venous thromboembolism prophylaxis and treatment. Despite their widespread application, heparin-induced skin lesions are probably under-reported and under-estimated. We present a case report of a primigravida treated with low molecular weight heparin for deep vein thrombosis, who developed a delayed-type hypersensitivity reaction to enoxaparin, tinzaparin and dalteparin. As the patient was pregnant, treatment options were restricted. Tolerance was achieved with dalteparin with adjuvant administration of prednisolone. An attempt to decrease prednisolone dose triggered delayed-type hypersensitivity reaction recurrence that was solved by keeping the initial prednisolone prescription. To the best of our knowledge, there are no described cases using this approach. In cases of delayed-type hypersensitivity reaction to low molecular weight heparin during pregnancy our case suggests that switching low molecular weight heparin and adjuvant administration of prednisolone can be an option.
PubMed: 38074199
DOI: 10.1177/1753495X221102709 -
Journal of Chromatography. A Dec 2023Matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a novel soft ionization bio-mass spectrometry technology emerging in the... (Review)
Review
Matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a novel soft ionization bio-mass spectrometry technology emerging in the 1980s, which can realize rapid detection of non-volatile, highly polar, and thermally unstable macromolecules. However, the analysis of small molecular compounds has been a major problem for MALDI-TOF MS all the time. In the MALDI analysis process based on traditional matrices, large numbers of interference peaks in the low molecular weight area and "sweet spots" phenomenon are produced, so the detection method needs to be further optimized. The promotion of matrix means the improvement of MALDI performance. In recent years, many new nanomaterial-based matrices have been successfully applied to the analysis of small molecular compounds, which makes MALDI applicable to a wider range of detection and useful in more fields such as pharmacy and environmental science. In this paper, the newly developed MALDI matrix categories in recent years are reviewed initially. Meanwhile, the potential applications, advantages and disadvantages of various matrices are analyzed. Finally, the future development prospects of nanomaterial-based matrices are also prospected.
Topics: Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Nanostructures; Molecular Weight; Lasers
PubMed: 37944434
DOI: 10.1016/j.chroma.2023.464493 -
Food Chemistry Nov 2023Low-molecular-weight-galactomannan (LMW-GM) is an edible polysaccharide with various biological activities. However, it is used in the field of neuroprotection. In this... (Comparative Study)
Comparative Study
Low-molecular-weight-galactomannan (LMW-GM) is an edible polysaccharide with various biological activities. However, it is used in the field of neuroprotection. In this study, two types of LMW-GMs from Sesbania cannabina were obtained by gluconic acid extraction (GA-LMW-GM) and enzymatic hydrolysis (GMOS). The structure of GA-LMW-GM and GMOS were identified using different nuclear magnetic resonance (NMR) techniques. The antioxidant and neuroprotective activities of GA-LMW-GM and GMOS were evaluated in vitro/vivo. The results showed that both GA-LMW-GM and GMOS possess good free radicals scavenging ability in vitro with IC values of 1.9 mg/mL and 4.9 mg/mL for 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals 2.8 mg/mL and 4.4 mg/mL for O radicals, respectively. However, GA-LMW-GM was more effective at scavenging reactive oxygen species (ROS) in vivo and protecting the fundamental growth (with a recovery capability of 62.5%) and locomotor functions (with recovery capability of 193.7%) of zebrafish with neurological damage induced by Bisphenol AF.
Topics: Animals; Antioxidants; Molecular Weight; Neuroprotection; Sesbania; Zebrafish
PubMed: 37364317
DOI: 10.1016/j.foodchem.2023.136642 -
Journal of Microbiology and... Jan 2024Low molecular weight collagen peptide (LMWCP) is a collagen hydrolysate derived from fish. We investigated the effects of LMWCP on hair growth using human dermal papilla...
Low molecular weight collagen peptide (LMWCP) is a collagen hydrolysate derived from fish. We investigated the effects of LMWCP on hair growth using human dermal papilla cells (hDPCs), human hair follicles (hHFs), patch assay, and telogenic C57BL/6 mice, while also examining the underlying mechanisms of its action. LMWCP promoted proliferation and mitochondrial potential, and the secretion of hair growth-related factors, such as EGF, HB-EGF, FGF-4, and FGF-6 in hDPCs. Patch assay showed that LMWCP increased the neogeneration of new HFs in a dose-dependent manner. This result correlated with an increase in the expression of dermal papilla (DP) signature genes such as, , , , and . LMWCP upregulated phosphorylation of glycogen synthase kinase-3β (GSK-3β) and β-catenin, and nuclear translocation of β-catenin, and it increased the expression of Wnt3a, LEF1, VEGF, ALP, and β-catenin. LMWCP promoted the growth of hHFs and increased the expression of β-catenin and VEGF. Oral administration of LMWCP to mice significantly stimulated hair growth. The expression of Wnt3a, β-catenin, PCNA, Cyclin D1, and VEGF was also elevated in the back skin of the mice. Furthermore, LMWCP increased the expression of cytokeratin and Keratin Type I and II. Collectively, these findings demonstrate that LMWCP has the potential to increase hair growth via activating the Wnt/β-catenin signaling pathway.
Topics: Mice; Humans; Animals; Wnt Signaling Pathway; Glycogen Synthase Kinase 3 beta; beta Catenin; Molecular Weight; Vascular Endothelial Growth Factor A; Cells, Cultured; Mice, Inbred C57BL; Hair Follicle; Hair; Cell Proliferation
PubMed: 37830229
DOI: 10.4014/jmb.2308.08013 -
Mutation Research 2024Inhaled anesthetics, such as isoflurane, may cause side effects, including short-term immunosuppression and DNA damage. In contrast, low molecular weight fucoidan (LMF),...
Inhaled anesthetics, such as isoflurane, may cause side effects, including short-term immunosuppression and DNA damage. In contrast, low molecular weight fucoidan (LMF), derived from brown seaweed, exhibits promising immunomodulatory effects. In this study, we determined the effect of isoflurane on telomeres and examined the potential of LMF to ameliorate the harmful effects of isoflurane. Male Lewis rats, the mouse lymphoma cell line YAC-1, and the human nature killer cell line NK-92 MI were exposed to isoflurane. The relative telomere length (T/S) ratio and mRNA expression were determined by quantitative PCR. The viability assay was used to assess cell viability. In vivo, 2% isoflurane exposure, which is a clinically relevant concentration, reduced telomere length, and correlated with exposure frequency and duration. Isoflurane concentrations above 2% shortened YAC-1 telomeres, with minimal impact on cell viability. LMF pre-treatment enhanced NK-92 MI cell survival resulting from isoflurane exposure and exerted superior telomere protection compared with LMF post-treatment. Furthermore, adding LMF during isoflurane exposure resulted in a significant increase in IFN-γ, TNF-α, and IL-10 mRNA compared with the untreated group. LMF protected against isoflurane-induced telomere shortening, enhanced NK cell viability, and modulated cytokine expression, thus mitigating postoperative immune suppression and risk of tumor metastasis.
Topics: Animals; Polysaccharides; Isoflurane; Mice; Male; Humans; Rats; Killer Cells, Natural; Anesthetics, Inhalation; Cell Survival; Telomere; Rats, Inbred Lew; Molecular Weight; Cell Line, Tumor; Telomere Homeostasis
PubMed: 38603928
DOI: 10.1016/j.mrfmmm.2024.111857 -
Journal of Pharmaceutical Sciences Oct 2023Microbial transglutaminase (mTG) has become a powerful tool for manufacturing antibody-drug conjugates (ADCs). It enables site-specific conjugation by catalyzing...
Microbial transglutaminase (mTG) has become a powerful tool for manufacturing antibody-drug conjugates (ADCs). It enables site-specific conjugation by catalyzing formation of stable isopeptide bond between glutamine (Q) side chain and primary amine. However, the downstream impact of mTG-mediated conjugation on ADC product quality, especially on high molecular weight (HMW) size variant formation has not been studied in a systematic manner. This study investigates the mechanisms underlying the formation of HMW size variants in mTG-mediated ADCs using size exclusion chromatography (SEC) and liquid chromatography-mass spectrometry (LC-MS). Our findings revealed that the mTG-mediated glutamine and lysine (K) crosslinking is the primary source of the increased level of HMW size variants in the ADCs. In the study, two monoclonal antibodies (mAbs) with glutamine engineered for site-specific conjugation were used as model systems. Based on the LC-MS analysis, a single lysine (K56) in the heavy chain (HC) was identified as the major Q-K crosslinking site in one of the two mAbs. The HC C-terminal K was observed to crosslink to the target Q in both mAbs. Quantitative correlation was established between the percentage of HMW size variants determined by SEC and the percentage of crosslinked peptides quantified by MS peptide mapping. Importantly, it was demonstrated that the level of HMW size variants in the second ADC was substantially reduced by the complete removal of HC C-terminal K before conjugation. The current work demonstrates that crosslinking and other side reactions during mTG-mediated conjugation needs to be carefully monitored and controlled to ensure process consistency and high product quality of the final ADC drug product.
Topics: Immunoconjugates; Transglutaminases; Molecular Weight; Lysine; Glutamine; Antibodies, Monoclonal
PubMed: 37586591
DOI: 10.1016/j.xphs.2023.08.006 -
Chemosphere Oct 2023Atmospheric brown carbon (BrC) contain amounts of organic species, but their molecular weight (MW) distributions is still poorly understood. This study applied...
Investigating the molecular weight distribution of atmospheric water-soluble brown carbon using high-performance size exclusion chromatography coupled with diode array and fluorescence detectors.
Atmospheric brown carbon (BrC) contain amounts of organic species, but their molecular weight (MW) distributions is still poorly understood. This study applied high-performance size exclusion chromatography (HPSEC) coupled with a diode array detector (DAD) and fluorescence detector (FLD) to characterize the MW distributions of typical chromophores and fluorophores within water-soluble BrC. The investigation focused on the spring season, encompassing both typical urban and rural aerosols. Our results showed that chromophores (at 254 and 365 nm), and humic-like and protein-like fluorophores identified by excitation-emission matrix parallel factor analysis (EEM-PARAFAC) within BrC were broadly distributed along the MW continuum (∼50-20,000 Da). This suggests that BrC mainly comprises complex chromophores and fluorophores with heterogeneous molecular sizes. High-MW (HMW, >1 kDa) species (66%-74%) dominated the chromophores at 254 and 365 nm. However, the latter chromophores were enriched with more HMW species. This result suggested that the HMW chromophores might contribute more to BrC absorption at longer wavelengths. The PARAFAC-derived fluorescent components also exhibited different MW distributions. Three humic-like substances (HULIS) were all dominated by HMW fractions (51%-74%), but protein-like fluorescent component (PLOM) enriched low-MW (LMW, <1 kDa) species (60%-66%). Furthermore, the molecular size (i.e., weight-averaged and number-averaged MW) and the ratios between HMW and LMW species decreased in the order highly-oxygenated HULIS > less-oxygenated HULIS > PLOM, indicating that the fluorophores with longer Em were generally related to larger MW. To our knowledge, this is the first report on the molecular size of individual fluorescent components within aerosol BrC. The results obtained here enhanced our knowledge of heterogeneous composition, complex physicochemical properties, and potential atmospheric fates of aerosol BrC.
Topics: Carbon; Molecular Weight; Water; Chromatography, Gel; Humic Substances; Fluorescent Dyes; Aerosols; Environmental Monitoring
PubMed: 37454992
DOI: 10.1016/j.chemosphere.2023.139517 -
Cellular and Molecular Life Sciences :... Oct 2023Although the pro-tumorigenic functions of hyaluronan (HA) are well documented there is limited information on the effects and targets of different molecular weight HA....
Although the pro-tumorigenic functions of hyaluronan (HA) are well documented there is limited information on the effects and targets of different molecular weight HA. Here, we investigated the effects of 27 kDa, 183 kDa and 1000 kDa HA on ES-2 ovarian cancer cells overexpressing the stem cell associated protein, Notch3. 1000 kDA HA promoted spheroid formation in ES-2 cells mixed with ES-2 overexpressing Notch3 (1:3). We report disabled-2 (DAB2) as a novel protein regulated by 1000 kDa HA and further investigated its role in ovarian cancer. DAB2 was downregulated in ovarian cancer compared to normal tissues but increased in metastatic ovarian tumors compared to primary tumors. High DAB2 expression was associated with poor patient outcome and positively correlated with HA synthesis enzyme HAS2, HA receptor CD44 and EMT and macrophage markers. Stromal DAB2 immunostaining was significantly increased in matched ovarian cancer tissues at relapse compared to diagnosis and associated with reduced survival. The proportion of DAB2 positive macrophages was significantly increased in metastatic ovarian cancer tissues compared to primary cancers. However, DAB2 overexpression significantly reduced invasion by both A2780 and OVCAR3 cells in vivo. Our research identifies a novel relationship between HA signalling, Notch3 and DAB2. We highlight a complex relationship of both pro-tumorigenic and tumor suppressive functions of DAB2 in ovarian cancer. Our findings highlight that DAB2 has a direct tumor suppressive role on ovarian cancer cells. The pro-tumorigenic role of DAB2 may be mediated by tumour associated macrophages and requires further investigation.
Topics: Female; Humans; Apoptosis; Cell Line, Tumor; Hyaluronan Receptors; Hyaluronic Acid; Molecular Weight; Ovarian Neoplasms; Tumor Suppressor Proteins
PubMed: 37815603
DOI: 10.1007/s00018-023-04972-9