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Microorganisms Apr 2024Representatives of the genus are widely used as probiotics to modulate the gut microbiome and alleviate various health conditions. The action mechanisms of probiotics...
Representatives of the genus are widely used as probiotics to modulate the gut microbiome and alleviate various health conditions. The action mechanisms of probiotics rely on their direct effect on the gut microbiota and the local and systemic effect of its metabolites. The main purpose of this animal experiment was to assess the biosafety of the strain BIOCC1719. Additional aims were to characterise the influence of the strain on the intestinal microbiota and the effect on several health parameters of the host during 15- and 30-day oral administration of the strain to mice. The strain altered the gut microbial community, thereby altering luminal short-chain fatty acid metabolism, resulting in a shift in the proportions of acetic, butyric, and propionic acids in the faeces and serum of the test group mice. Targeted metabolic profiling of serum revealed the possible ability of the strain to positively affect the hosts' amino acids and bile acids metabolism, as the cholic acid, deoxycholic acid, aspartate, and glutamate concentration were significantly higher in the test group. The tendency to increase anti-inflammatory polyamines (spermidine, putrescine) and neuroprotective 3-indolepropionic acid metabolism and to lower uremic toxins (P-cresol-SO, indoxyl-SO) was registered. Thus, BIOCC1719 may exert health-promoting effects on the host through modulation of the gut microbiome and the host metabolome via inducing the production of health-promoting bioactive compounds. The health effects of the strain need to be confirmed in clinical trials with human volunteers.
PubMed: 38674784
DOI: 10.3390/microorganisms12040840 -
Biomolecules Apr 2024Polyamines are polycations derived from amino acids that play an important role in proliferation and growth in almost all living cells. In (the pneumococcus),...
Polyamines are polycations derived from amino acids that play an important role in proliferation and growth in almost all living cells. In (the pneumococcus), modulation of polyamine metabolism not only plays an important regulatory role in central metabolism, but also impacts virulence factors such as the capsule and stress responses that affect survival in the host. However, functional annotation of enzymes from the polyamine biosynthesis pathways in the pneumococcus is based predominantly on computational prediction. In this study, we cloned SP_0166, predicted to be a pyridoxal-dependent decarboxylase, from the Orn/Lys/Arg family pathway in TIGR4 and expressed and purified the recombinant protein. We performed biochemical characterization of the recombinant SP_0166 and confirmed the substrate specificity. For polyamine analysis, we developed a simultaneous quantitative method using hydrophilic interaction liquid chromatography (HILIC)-based liquid chromatography-tandem mass spectrometry (LC-MS/MS) without derivatization. SP_0166 has apparent , , and / values of 11.3 mM, 715,053 min, and 63,218 min mM, respectively, with arginine as a substrate at pH 7.5. We carried out inhibition studies of SP_0166 enzymatic activity with arginine as a substrate using chemical inhibitors DFMO and DFMA. DFMO is an irreversible inhibitor of ornithine decarboxylase activity, while DFMA inhibits arginine decarboxylase activity. Our findings confirm that SP_0166 is inhibited by DFMA and DFMO, impacting agmatine production. The use of arginine as a substrate revealed that the synthesis of putrescine by agmatinase and -carbamoylputrescine by agmatine deiminase were both affected and inhibited by DFMA. This study provides experimental validation that SP_0166 is an arginine decarboxylase in pneumococci.
Topics: Carboxy-Lyases; Tandem Mass Spectrometry; Streptococcus pneumoniae; Chromatography, High Pressure Liquid; Substrate Specificity; Bacterial Proteins; Recombinant Proteins; Polyamines; Kinetics
PubMed: 38672479
DOI: 10.3390/biom14040463 -
Journal of Agricultural and Food... May 2024Drought stress has become the primary severe threat to global agriculture production, including medicinal plants. Plant growth-promoting bacteria (PGPB) and...
Drought stress has become the primary severe threat to global agriculture production, including medicinal plants. Plant growth-promoting bacteria (PGPB) and environmentally friendly element silicon (Si) have emerged as effective methods in alleviating drought stress in various plants. Here, the effects of the plant endophytic G5 interaction with Si on regulating nitrogen absorption, assimilation, and metabolism pathways were investigated in the morphophysiological and gene attributes of exposed to drought. Results showed that G5+Si application improved nitrogen absorption and assimilation by increasing the available nitrogen content in the soil, further improving the nitrogen utilization efficiency. Then, G5+Si triggered the accumulation of the major adjustment substances proline, γ-aminobutyric acid, putrescine, and chlorophyll, which played an important role in contributing to maintaining balance and energy supply in exposed to drought. These findings will provide new ideas for the combined application of PGPR and Si on both soil and plant systems in a drought habitat.
Topics: Droughts; Nitrogen; Silicon; Endophytes; Glycyrrhiza uralensis; Bacillus; Stress, Physiological; Chlorophyll; Soil; Plant Roots
PubMed: 38661009
DOI: 10.1021/acs.jafc.4c00282 -
Breast Cancer Research : BCR Apr 2024Basal-like breast cancer (BLBC) is the most aggressive subtype of breast cancer due to its aggressive characteristics and lack of effective therapeutics. However, the...
BACKGROUND
Basal-like breast cancer (BLBC) is the most aggressive subtype of breast cancer due to its aggressive characteristics and lack of effective therapeutics. However, the mechanism underlying its aggressiveness remains largely unclear. S-adenosylmethionine decarboxylase proenzyme (AMD1) overexpression occurs specifically in BLBC. Here, we explored the potential molecular mechanisms and functions of AMD1 promoting the aggressiveness of BLBC.
METHODS
The potential effects of AMD1 on breast cancer cells were tested by western blotting, colony formation, cell proliferation assay, migration and invasion assay. The spermidine level was determined by high performance liquid chromatography. The methylation status of CpG sites within the AMD1 promoter was evaluated by bisulfite sequencing PCR. We elucidated the relationship between AMD1 and Sox10 by ChIP assays and quantitative real-time PCR. The effect of AMD1 expression on breast cancer cells was evaluated by in vitro and in vivo tumorigenesis model.
RESULTS
In this study, we showed that AMD1 expression was remarkably elevated in BLBC. AMD1 copy number amplification, hypomethylation of AMD1 promoter and transcription activity of Sox10 contributed to the overexpression of AMD1 in BLBC. AMD1 overexpression enhanced spermidine production, which enhanced eIF5A hypusination, activating translation of TCF4 with multiple conserved Pro-Pro motifs. Our studies showed that AMD1-mediated metabolic system of polyamine in BLBC cells promoted tumor cell proliferation and tumor growth. Clinically, elevated expression of AMD1 was correlated with high grade, metastasis and poor survival, indicating poor prognosis of breast cancer patients.
CONCLUSION
Our work reveals the critical association of AMD1-mediated spermidine-eIF5A hypusination-TCF4 axis with BLBC aggressiveness, indicating potential prognostic indicators and therapeutic targets for BLBC.
Topics: Humans; Female; Breast Neoplasms; Eukaryotic Translation Initiation Factor 5A; Peptide Initiation Factors; Mice; Animals; Gene Expression Regulation, Neoplastic; Cell Proliferation; Spermidine; RNA-Binding Proteins; Transcription Factor 4; Cell Line, Tumor; Promoter Regions, Genetic; Adenosylmethionine Decarboxylase; Cell Movement; DNA Methylation; Prognosis; SOXE Transcription Factors; Lysine
PubMed: 38654332
DOI: 10.1186/s13058-024-01825-6 -
Frontiers in Plant Science 2024This study was envisaged to investigate the physiological reasons affecting the embryo development and abortion of seedless grapes on the basis of the previous embryo...
This study was envisaged to investigate the physiological reasons affecting the embryo development and abortion of seedless grapes on the basis of the previous embryo rescue breeding techniques of seedless grapes. Specifically, the relationship between the embryo rescue breeding of seedless grapes and the change of polyamine content was evaluated, in order to provide hybrid germplasm in the breeding of new seedless grape cultivars. Four ovules of 4 naturally pollinated Eurasian seedless grape cultivars, including 'Thompson Seedless' grape (hereinafter referred to as 'Seedless White' grape), 'Flame Seedless' grape, 'Heshi Seedless' grape and 'Ruby Seedless' grape were employed for the study. Changes in the endogenous polyamine content, exogenous polyamine content, and the suitable combination of exogenous polyamines in the seedless grape berries and isolated ovules were determined during the best embryo rescue period. Furthermore, the effect of different exogenous polyamine contents on the germination and seedling rate of different seedless grape embryos was analyzed. In the best embryo rescue period, the number of ovules had different effects on the content of polyamines. For seedless grape cultivars with 4 ovules, a high content of polyamines was found to be more beneficial in the embryonic development. The existence of embryos had different effects on the development of embryos. In the ovules with embryo, an increase in the content of polyamine was beneficial to the growth and development of the ovule. Different ratios of exogenous polyamines had varying effects on the embryonic development. Putrescine (Put) exhibited the greatest effect on the embryonic development. Further, correlation analysis showed that different combinations of exogenous polyamines had varying effects on the embryonic development. A maximal ovule development was observed in the combination of exogenous polyamines of putrescine2+spermidine2+spermine1. For maximal embryo germination and seeding formation, the optimal combination was putrescine2+spermidine2+spermine2. Irrespective to the number of ovules or the existence of embryos, the results indicated that a high content of endogenous polyamines promoted the growth and development of embryos. The embryo rescue efficiency of different exogenous polyamines was different, and the appropriate combination of exogenous polyamines was beneficial to the growth and development of ovules, with a high development rate of the ovule and seedling.
PubMed: 38650703
DOI: 10.3389/fpls.2024.1362989 -
Pharmacology, Biochemistry, and Behavior Jul 2024Memory consolidation is associated with the regulation of protein kinases, which impact synaptic functions and promote synaptogenesis. The administration of spermidine...
Memory consolidation is associated with the regulation of protein kinases, which impact synaptic functions and promote synaptogenesis. The administration of spermidine (SPD) has been shown to modulate major protein kinases associated with memory improvement, including the Ca-dependent protein kinase (PKC) and cAMP-dependent protein kinase (PKA), key players in the cAMP response element-binding protein (CREB) activation. Nevertheless, the initial mechanism underlying SPD-mediated memory consolidation remains unknown, as we hypothesize a potential involvement of the memory consolidation precursor, Ca2+/calmodulin-dependent protein kinase II-α (CaMKIIα), in this process. Based on this, our study aimed to investigate potential interactions among PKC, PKA, and CREB activation, mediated by CaMKIIα activation, in order to elucidate the SPD memory consolidation pathway. Our findings suggest that the post-training administration of the CaMKII inhibitor, KN-62 (0.25 nmol, intrahippocampal), prevented the memory enhancement induced by SPD (0.2 nmol, intrahippocampal) in the inhibitory avoidance task. Through western immunoblotting, we observed that phosphorylation of CaMKIIα in the hippocampus was facilitated 15 min after intrahippocampal SPD administration, resulting in the activation of PKA and CREB, 180 min after infusion, suggesting a possible sequential mechanism, since SPD with KN-62 infusion leads to a downregulation in CaMKIIα/PKA/CREB pathway. However, KN-62 does not alter the memory-facilitating effect of SPD on PKC, possibly demonstrating a parallel cascade in memory acquisition via PKA, without modulating CAMKIIα. These results suggest that memory enhancement induced by SPD administration involves crosstalk between CaMKIIα and PKA/CREB, with no PKC interaction.
Topics: Animals; Calcium-Calmodulin-Dependent Protein Kinase Type 2; Cyclic AMP-Dependent Protein Kinases; Rats; Spermidine; Male; Cyclic AMP Response Element-Binding Protein; Memory; Rats, Wistar; Signal Transduction; Hippocampus; Phosphorylation; Sulfonamides; Benzylamines; Avoidance Learning; Protein Kinase C; 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine
PubMed: 38648866
DOI: 10.1016/j.pbb.2024.173774 -
Research Square Apr 2024Latent tuberculosis infection (LTBI) is common in people living with HIV (PLHIV) in high TB burden settings. Active TB is associated with specific stool taxa; however,...
BACKGROUND
Latent tuberculosis infection (LTBI) is common in people living with HIV (PLHIV) in high TB burden settings. Active TB is associated with specific stool taxa; however, little is known about the stool microbiota and LTBI, including in PLHIV.
METHOD
Within a parent study that recruited adult females with HIV from Cape Town, South Africa into predefined age categories (18-25, 35-60 years), we characterised the stool microbiota of those with [interferon-γ release assay (IGRA)- and tuberculin skin test (TST)-positive] or without (IGRA- and TST- negative) LTBI (n=25 per group). 16S rRNA DNA sequences were analysed using QIIME2, Dirichlet Multinomial Mixtures, DESeq2 and PICRUSt2.
RESULTS
No α- or β-diversity differences occurred by LTBI status; however, LTBI-positives were depleted. Inferred metagenome data showed LTBI-negative-enriched pathways included several involved in methylglyoxal degradation, L-arginine, putrescine, 4-aminobutanoate degradation and L-arginine and ornithine degradation. Stool from LTBI-positives demonstrated differential taxa abundance based on a quantitative response to antigen stimulation (depletion associated with higher IGRA or TST responses, respectively). In LTBI-positives, older people had different β-diversities than younger people whereas, in LTBI-negatives, no differences occurred across age groups.
CONCLUSION
Amongst female PLHIV, those with LTBI had, vs. those without LTBI, Gemmiger-enriched, which are producers of short chain fatty acids. Taxonomic differences amongst people with LTBI occurred according to quantitative response to antigen stimulation and age. These data enhance our understanding of the microbiome's potential role in LTBI.
PubMed: 38645218
DOI: 10.21203/rs.3.rs-4182285/v1 -
DAB-APT: a Fluorescence-Based Assay for Determining Aminopropyl Transferase Activity and Inhibition.BioRxiv : the Preprint Server For... Apr 2024Polyamines are polycationic molecules that are crucial in a wide array of cellular functions. Their biosynthesis is mediated by aminopropyl transferases (APTs),...
Polyamines are polycationic molecules that are crucial in a wide array of cellular functions. Their biosynthesis is mediated by aminopropyl transferases (APTs), promising targets in antimicrobial, antineoplastic and antineurodegenerative therapies. A major limitation, however, is the lack of high-throughput assays to measure their activity. We developed the first fluorescence-based assay, DAB-APT, for measurement of APT activity using 1,2-diacetyl benzene, which forms fluorescent conjugates with putrescine, spermidine and spermine with fluorescence intensity increasing with increasing carbon chain length. The assay has been validated using APT enzymes from and and is suitable for high-throughput screening of large chemical libraries. Given the importance of APTs in infectious diseases, cancer and neurobiology, our DAB-APT assay has broad applications, holding promise for advancing research and drug discovery efforts.
PubMed: 38645036
DOI: 10.1101/2024.04.09.588734 -
Physiologia Plantarum 2024The flower bud differentiation plays a crucial role in cherry yield and quality. In a preliminary study, we revealed the promotion of spermidine (Spd) in bud...
The flower bud differentiation plays a crucial role in cherry yield and quality. In a preliminary study, we revealed the promotion of spermidine (Spd) in bud differentiation and quality. However, the molecular mechanism underlying Spd regulating cherry bud differentiation remains unclear. To address this research gap, we cloned CpSPDS2, a gene that encodes Spd synthase and is highly expressed in whole flowers and pistils of the Chinese cherry (cv. 'Manaohong'). Furthermore, an overexpression vector with this gene was constructed to transform tobacco plants. The findings demonstrated that transgenic lines exhibited higher Spd content, an earlier flowering time by 6 d, and more lateral buds and flowers than wild-type lines. Additionally, yeast one-hybrid assays and two-luciferase experiments confirmed that the R2R3-MYB transcription factor (CpMYB44) directly binds to and activates the CpSPDS2 promoter transcription. It is indicated that CpMYB44 promotes Spd accumulation via regulating CpSPDS2 expression, thus accelerating the flower growth. This research provides a basis for resolving the molecular mechanism of CpSPDS2 involved in cherry bud differentiation.
Topics: Spermidine; Nicotiana; Plant Proteins; Gene Expression Regulation, Plant; Transcription Factors; Prunus; Flowers
PubMed: 38629194
DOI: 10.1111/ppl.14300 -
Food Science & Nutrition Apr 2024This study aimed at investigating the quality attributes of Turkish fermented sausage (sucuk) incorporated with bay leaf extract obtained as a natural antioxidant and a...
This study aimed at investigating the quality attributes of Turkish fermented sausage (sucuk) incorporated with bay leaf extract obtained as a natural antioxidant and a source of phytochemicals. Five different bay leaf extracts were obtained with distilled water and 60%, 70%, 80%, and 90% ethanol. The total phenolic contents and antioxidant activity values indicated that ultrasound-assisted 70% ethanol extract was the most suitable extract. Furthermore, five groups of sucuks were manufactured with the addition of bay leaf extract (1, 5, and 10 mL/kg), ascorbic acid (500 mg/kg), and control. The extracts were produced similar pH values when compared to control and ascorbic acid samples. The treatments had no significant effect on moisture contents of sucuks. Bay leaf extracts produced comparable color, texture profile analysis, and TBARS values to control and ascorbic acid samples. Biogenic amine contents (mg/kg dry weight) of sucuks including tryptamine (6.43-30.66), 2-phenylethylamine (2.24-32.04), putrescine (2.19-7.98), cadaverine (3.28-12.21), histamine (7.01-11.38), tyramine (3.27-71.07), spermidine (4.44-8.01), and spermine (53.96-68.25) were mostly within the lower ranges typically associated with sucuk. However, the lowest cadaverine values observed at the end of storage in the bay leaf extract added samples indicated that bay leaf extract might be effective in decreasing cadaverine values during storage. The addition of bay leaf extract caused similar sensory attributes to the control and ascorbic acid samples. This study revealed that Turkish fermented sucuks could be effectively incorporated with bay leaf extracts without a negative effect on the quality attributes or consumer acceptability.
PubMed: 38628223
DOI: 10.1002/fsn3.3929