Sugar Residues Research Articles

Collected Thoughts on Mycobacterial Lipoarabinomannan, a Cell Envelope Lipoglycan.

The presence of lipoarabinomannan (LAM) in the Mycobacterium tuberculosis (Mtb) cell envelope was first reported close to 100 years ago. Since then, numerous studies have been dedicated to the isolation, purification, structural definition, and elucidation of the biological properties of Mtb LAM. In this review, we present a brief historical perspective on the discovery of Mtb LAM and the herculean efforts devoted to structurally characterizing the molecule because of its unique structural and biological features. The significance of LAM remains high to this date, mainly due to its distinct immunological properties in conjunction with its role as a biomarker for diagnostic tests due to its identification in urine, and thus can serve as a point-of-care diagnostic test for tuberculosis (TB). In recent decades, LAM has been thoroughly studied and massive amounts of information on this intriguing molecule are now available. In this review, we give the readers a historical perspective and an update on the current knowledge of LAM with information on the inherent carbohydrate composition, which is unique due to the often puzzling sugar residues that are specifically found on LAM. We then guide the readers through the complex and myriad immunological outcomes, which are strictly dependent on LAM's chemical structure. Furthermore, we present issues that remain unresolved and represent the immediate future of LAM research. Addressing the chemistry, functions, and roles of LAM will lead to innovative ways to manipulate the processes that involve this controversial and fascinating biomolecule.

Effects of oyster shells on maturity and calcium activation in organic solid waste compost

With the rapid development of aquaculture, the production of oyster shells has surged, posing a potential threat to the environment. While oyster shell powder is widely recognized for its inherent alkalinity and rich calcium carbonate content, making it a superior soil conditioner, its role in organic solid waste composting remains underexplored. To investigate the effects of varying concentrations of oyster shell powder on compost maturation and calcium activation, this study employed thermophilic co-composting with acidic sugar residue and bean pulp, incorporating 0% (control), 10% (T1), 20% (T2), 30% (T3), and 40% (T4) oyster shell powder. Findings revealed that appropriate proportions of oyster shell powder significantly enhance temperature stability during composting and elevate maturation levels, notably reducing ammonia emissions between 62.5% and 76.7%. Intriguingly, the calcium in the oyster shell powder was significantly activated during composting, with the 40% addition group achieving the highest calcium activation rate of 48.5%. In summation, the inclusion of oyster shell powder not only optimizes the composting process but also efficiently activates the calcium, resulting in an alkaline organic-inorganic composite soil conditioner with high exchangeable calcium content. This research holds significant implications for promoting the high-value utilization of oyster shells.

Sulfur-containing spiroketals from Breynia disticha and evaluations of their anti-inflammatory effect.

Breynia spp. are a key source of sulfur-containing spiroketal glycosides with potential anti-inflammatory activity. In this study, three new sulfur-containing spiroketals - breynin J (1), epibreynin J (2), and probreynogenin (3) - along with four known compounds - probreynin I (4), phyllaemblic acid (5), breynin B (6), and epibreynin B (7) - were isolated from the roots of Breynia disticha. The structures of compounds 1-7 were elucidated by extensive 1D and 2D NMR spectroscopic analyses, including 1D total correlation spectroscopy (TOCSY), HSQC, HMBC, double quantum-filtered (DQF)-COSY, heteronuclear two-bond correlation (H2BC), and HSQC-TOCSY experiments, as well as high-resolution electrospray ionization HRESIMS analysis, and quantum chemical electronic CD calculations. Furthermore, the absolute configurations of sugar residues were determined by derivatization of the hydrolysates with ʟ-cysteine methyl ester and o-tolyl isothiocyanate followed by HPLC analysis. The anti-inflammatory effects of the isolated compounds were evaluated based on the mRNA levels of proinflammatory cytokines in lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophage cells. Compounds 1, 2, 6, and 7 inhibited the increase in interleukin (IL)-1β and IL-6 mRNA levels stimulated by LPS. Moreover, the most potent compound 7 was found to significantly inhibit the production of IL-1β and IL-6 proteins, as revealed by the analysis of culture supernatants.

A novel polysaccharide from the fruits of Cudrania tricuspidata and its antioxidant and alcohol dehydrogenase activating ability

In the present study, polysaccharides from the fruits of Cudrania tricuspidata were purified hierarchically using DEAE-52 cellulose columns and Sephadex S-100 gel columns. Two fractions were obtained and named ZGs-1 and ZGs-2. The in vitro antioxidant and alcohol dehydrogenase activities of ZGs-1 and ZGs-2 were also analyzed. Two grades named ZGs-1 and ZGs-2 were obtained with molecular weights of 131.521 and 94.551 KDa, respectively, and both consisted of arabinose, galactose, glucose and small amounts of xylose and rhamnose. The structural resolution of these two levels by FT-IR, methylation and NMR showed that the major sugar residue types of ZGs-1 are: →3)-β-D-Galp-(1→, α-L-Araf-(1→, →3,6)-β-D-Galp-(1→, β-D-Xylp-(1→, →5)-α-L-Araf-(1→, →3,5)-α-L-Araf-(1→, →4)-α-D-Glc-(1→. The main sugar residue types of ZGs-2 are: →6)-β-D-Galp-(1→, →5)-α-L-Araf-(1→, →3,6)-β-D-Galp-(1→, →3,5)-α-L-Araf-(1→, →4)-β-D-Glc-(1→, α-L-Araf-(1→. ZGs-1 showed weak antioxidant activity. ZGs-2 showed good scavenging effects on DPPH and hydroxyl radicals, as well as on iron ion reduction ability, and the activation rate of alcohol dehydrogenase reached 92.15%, indicating that the novel polysaccharide ZGs-2 is a potential antioxidant and hepatoprotective agent.

Type I Lipoteichoic Acid (LTA) Purification by Hydrophobic Interaction Chromatography and Structural Analysis by 2D Nuclear Magnetic Resonance (NMR) Spectroscopy.

Type I lipoteichoic acid (LTA) is a glycerolphosphate polymer found in the cell envelope of diverse Gram-positive bacteria. The glycerolphosphate backbone is often further decorated with D-alanine and/or sugar residues. Here, we provide details of a 1-butanol extraction and purification method of type I LTA by hydrophobic interaction chromatography. The protocol has been adapted from methods originally described by Fischer et al. (Eur J Biochem 133:523-530, 1983) and further optimized by Morath et al. (J Exp Med 193:393-397, 2001). We also present information on a 2D nuclear magnetic resonance (NMR) analysis method to gain chemical and structural information of the purified LTA material.

A Neutral Polysaccharide from Spores of Ophiocordyceps gracilis Regulates Oxidative Stress via NRF2/FNIP1 Pathway.

Ophiocordyceps gracilis (O. gracilis) is a parasitic fungus used in traditional Chinese medicine and functional foods. In this study, a neutral heteropolysaccharide (GSP-1a) was isolated from spores of O. gracilis, and its structure and antioxidant capacities were investigated. GSP-1a was found to have a molecular weight of 72.8 kDa and primarily consisted of mannose (42.28%), galactose (35.7%), and glucose (22.02%). The backbone of GSP-1a was composed of various sugar residues, including →6)-α-D-Manp-(1→, →2,6)-α-D-Manp-(1→, →2,4,6)-α-D-Manp-(1→, →6)-α-D-Glcp-(1→, and →3,6)-α-D-Glcp-(1→, with some branches consisting of →6)-α-D-Manp-(1→ and α-D-Gal-(1→. In vitro, antioxidant activity assays demonstrated that GSP-1a exhibited scavenging effects on hydroxyl radical (•OH), 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid radical cation (ABTS•+), and 2,2-diphenyl-1-picrylhydrazyl radical (DPPH•). Moreover, GSP-1a was found to alleviate H2O2-induced oxidative stress in HepG2 cells by reducing the levels of reactive oxygen species (ROS) and malondialdehyde (MDA), while enhancing the activities of superoxide dismutase (SOD). Furthermore, GSP-1a upregulated the mRNA expression of antioxidant enzymes such as Ho-1, Gclm, and Nqo1, and regulated the NRF2/KEAP1 and FNIP1/FEM1B pathways. The findings elucidated the structural types of GSP-1a and provided a reliable theoretical basis for its usage as a natural antioxidant in functional foods or medicine.

Deacetylated Sialic Acid Sensitizes Lung and Colon Cancers to Novel Cucurbitacin-Inspired Estrone Epidermal Growth Factor Receptor (EGFR) Inhibitor Analogs.

Cancers utilize sugar residues such as sialic acids (Sia) to improve their ability to survive. Sia presents a variety of functional group alterations, including O-acetylation on the C6 hydroxylated tail. Previously, sialylation has been reported to suppress EGFR activation and increase cancer cell sensitivity to Tyrosine Kinase Inhibitors (TKIs). In this study, we report on the effect of deacetylated Sia on the activity of three novel EGFR-targeting Cucurbitacin-inspired estrone analogs (CIEAs), MMA 294, MMA 321, and MMA 320, in lung and colon cancer cells. Acetylation was modulated by the removal of Sialate O-Acetyltransferase, also known as CAS1 Domain-containing protein (CASD1) gene via CRISPR-Cas9 gene editing. Using a variety of cell-based approaches including MTT cell viability assay, flow cytometry, immunofluorescence assay and in-cell ELISA we observed that deacetylated Sia-expressing knockout cells (1.24-6.49 μM) were highly sensitive to all CIEAs compared with the control cells (8.82-20.97 μM). Apoptosis and varied stage cell cycle arrest (G0/G1 and G2/M) were elucidated as mechanistic modes of action of the CIEAs. Further studies implicated overexpression of CIEAs' cognate protein target, phosphorylated EGFR, in the chemosensitivity of the deacetylated Sia-expressing knockout cells. This observation correlated with significantly decreased levels of key downstream proteins (phosphorylated ERK and mTOR) of the EGFR pathway in knockout cells compared with controls when treated with CIEAs. Collectively, our findings indicate that Sia deacetylation renders lung and colon cancer cells susceptible to EGFR therapeutics and provide insights for future therapeutic interventions.

Selective targeting of lectins and their macropinocytosis in urothelial tumours: translation from in vitro to ex vivo

Urinary bladder cancer can be treated by intravesical application of therapeutic agents, but the specific targeting of cancer urothelial cells and the endocytotic pathways of the agents are not known. During carcinogenesis, the superficial urothelial cells exhibit changes in sugar residues on the apical plasma membranes. This can be exploited for selective targeting from the luminal side of the bladder. Here we show that the plant lectins Jacalin (from Artocarpus integrifolia), ACA (from Amaranthus caudatus) and DSA (from Datura stramonium) selectively bind to the apical plasma membrane of low- (RT4) and high-grade (T24) cancer urothelial cells in vitro and urothelial tumours ex vivo. The amount of lectin binding was significantly different between RT4 and T24 cells. Endocytosis of lectins was observed only in cancer urothelial cells and not in normal urothelial cells. Transmission electron microscopy analysis showed macropinosomes, endosome-like vesicles and multivesicular bodies filled with lectins in RT4 and T24 cells and also in cells of urothelial tumours ex vivo. Endocytosis of Jacalin and ACA in cancer cells was decreased in vitro after addition of inhibitor of macropinocytosis 5-(N-ethyl-N-isopropyl) amiloride (EIPA) and increased after stimulation of macropinocytosis with epidermal growth factor (EGF). Clathrin, caveolin and flotillin did not colocalise with lectins. These results confirm that the predominant mechanism of lectin endocytosis in cancer urothelial cells is macropinocytosis. Therefore, we propose that lectins in combination with conjugated therapeutic agents are promising tools for improved intravesical therapy by targeting cancer cells.

Study on the Biodegradation Process of D-Mannose Glycopolymers in Liquid Media and Soil.

Polymers derived from natural raw materials have become of great interest due to their increased biodegradable features and possible biocompatibility. Our group has successfully synthesized and characterized polymers derived from D-mannose oligomer (M), 2-hydroxy propyl acrylate (HPA), and methacrylate (HPMA) in different weight ratios. Their biodegradation was studied in liquid media with pure Proteus mirabilis inoculum for the samples with the most sugar residue, and the results show that the methacrylate derivative M_HPMA1 lost about 50% of its weight during incubation. SEM/EDX techniques were employed to display the modifications of the samples during the biodegradation process. The glycopolymers were buried in garden soil, and the experiment proved that more than 40% of the weight of the M_HPA1 sample was lost during biodegradation, while the other samples encountered an average of about 32% weight loss. The biodegradation profile was fitted against linear and polynomial mathematical models, which enabled an estimate of about a year for the total degradation of the D-mannose glycopolymers sample in soil.

BACTERIAL AMYLASE PRODUCTION BY THE L-LACTIC ACID-PRODUCING STRAIN USING CASSAVA STARCH SUPPLEMENTED WITH RICE BRAN

Amylase is a crucial starch-degrading enzyme, which is commercially available, prvides benefit to starchy industry to overcome viscosity problem that caused by using high starch concentration, and to supply sugar residues. This study aimed to investigate the production of amylase by the potential optical pure L-lactic acid-producing Lactococcus sp. SUT 513 for further development of the optical pure acid fermentation process from the cheap cassava starch. Rice bran, the main byproduct in the process of rice milling, was found to provide the maximum amylase production when supplemented with 4.0 g/L (w/v) to the medium containing cassava starch (20.0 g/L, w/v), compared to the cassava medium added either the expensive nitrogen source, tryptone (5.0 g/L) and yeast extract (3.0 g/L, w/v); or the cheap alternatives, soy ptotein (8.0 g/L) or urea (2.0 g/L) combined with yeast extract (3.0 g/L). The amylase activity obtained from the developed medium was approximately 3 times higher than the original medium. This bacterium gave the highest crude amylase activity of 4.848±0.237 U/mL in the suitable medium performed in a7.5-L bioreactor with 3.0-L working volume for the optimal cultivation time of only 18 h under a control pH of 7.0, temperature at 35°C, agitation at 150 rpm, and without aeration. The results reveal the high potential production of amylase by Lactococcus sp. SUT 513 using the cheap cassava starch medium supplemented with rice bran, providing an opportunity to develop both amylase and optically pure L-lactic acid production processes.

Structural, rheological and functional properties of ultrasonic treated xanthan gums

Xanthan gum can improve the freeze-thaw stability of frozen foods. However, the high viscosity and long hydration time of xanthan gum limits its application. In this study, ultrasound was employed to reduce the viscosity of xanthan gum, and the effect of ultrasound on its physicochemical, structural, and rheological properties was investigated using High-performance size-exclusion chromatography (HPSEC), ion chromatograph, methylation analysis, 1H NMR, rheometer, etc.. The application of ultrasonic-treated xanthan gum was evaluated in frozen dough bread. Results showed that the molecular weight of xanthan gum was reduced significantly by ultrasonication (from 3.0 × 107 Da to 1.4 × 106 Da), and the monosaccharide compositions and linkage patterns of sugar residues were altered. Results revealed that ultrasonication treatment mainly broke the molecular backbone at a lower intensity, then mainly broke the side chains with increasing intensity, which significantly reduced the apparent viscosity and viscoelastic properties of xanthan gum. The results of specific volume and hardness showed that the bread containing low molecular weight xanthan gum was of better quality. Overall, this work offers a theoretical foundation for broadening the application of xanthan gum and improving its performance in frozen dough.

AMYLOLYTIC POTENTIAL OF Arthrobacter sp. AND Bacillus aerius FROM WASTE DUMP-SITES OF TWO BAKERIES LOCATED WITHIN KADUNA METROPOLIS, NIGERIA

Bakery waste dumpsites are believed to harbour a variety of bacteria due to the presence of organic materials like flour, sugar, and other food residues. The conditions in these environments support the growth of bacteria with the ability to degrade starch into simple sugars because they produce amylases, an enzyme pivotal to the food and beverage production. As Amylases progressively gain relevance in several other starch-based processes, their availability becomes insufficient, and the need for amylases with unique and superior efficiency increases. The focus of this study was to isolate and characterize bacteria with amylase activity from the soil of bakery waste dumpsites to provide valuable insight into cheap sources of amylolytic bacteria and their potential biotechnological applications. The starch-agar medium was used to isolate and screen amylase-producing bacteria by streak plate method. Two isolates, A2 and A4, showing the most hydrolytic activities of 21 + 0.1 mm and 25 + 0.1 mm, respectively, were identified by their cultural and cellular morphologies, biochemical characteristics, 16S rRNA gene sequence, and nearest phylogenetic neighbour as Arthrobacter sp. and Bacillus aerius. Based on their culture and amylolytic temperatures, Arthrobacter sp. (37 0C) and Bacillus aerius (45 0C) tend to produce amylases that can be explored in low to moderate and moderate to torrid industrial processes, respectively.

Lectin histochemistry of posterior lingual glands of developing rats

The posterior lingual glands are classified as Weber and von Ebner glands. Glycans play an important role in salivary glands. Although the distribution of glycans can explain functional diversity and variation, there are many unknowns in the developing rat posterior lingual glands. The purpose of this study was to elucidate the relationship between the development and function of the posterior lingual gland in rats by histochemical analysis using lectins that bind to sugar residues. In adult rats, Arachis hypogaea (PNA), Glycine maximus (SBA), and Triticum vulgaris (WGA) were associated with serous cells and Dolichos biflorus (DBA) with mucous cells. In both Weber's and von Ebner's glands, all 4 lectins were bound to serous cells in early development, but as development progressed, DBA disappeared in serous cells and only the DBA remained in mucous cells. These results suggest that Galβ (1,3) > Galβ(1,4) > Gal, αGalNAc > αGal > βGalNAc, NeuAc > (GalNAc)2–3>>>GlcNAc, and GalNAcα(1,3) are present in the early stage of development, but that GalNAcα(1,3) disappear in serous cells and only GalNAcα(1,3) are localized in mucous cells after maturation. These results indicate that Weber glands function as serous glands in the early postnatal stage when von Ebner glands have not matured.

Effects of degradation on the physicochemical and antioxidant properties of carboxymethyl pachymaran

Poria cocos (Schw.) Wolf is a well-known edible and medicinal fungus. The polysaccharide in the sclerotium of P. cocos was extracted and prepared into carboxymethyl pachymaran (CMP). Three different degradation treatments including high temperature (HT), high pressure (HP) and gamma irradiation (GI) were used to process CMP. The changes in physicochemical properties and antioxidant activities of CMP were then comparatively investigated. We found that the molecular weights of HT-CMP, HP-CMP, and GI-CMP decreased from 787.9 kDa to 429.8, 569.5 and 6.0 kDa, respectively. Degradation treatments had no effect on the main chains of →3-β-D-Glcp-(1 → while changed the branched sugar residues. The polysaccharide chains of CMP were depolymerized after high pressure and gamma irradiation treatments. The three degradation methods improved the stability of CMP solution while decreased the thermal stability of CMP. In addition, we found that the GI-CMP with lowest molecular weight had the best antioxidant activity. Our results suggest that gamma irradiation treatment could degrade CMP as functional foods with strong antioxidant activity.

O-256 Evaluation of the fertility potential of semen using Proton NMR

Abstract Study question Is Proton NMR seminal plasma assessment a valid method for identifying biomarkers allowing to predict top-quality (TQ) embryo development? Summary answer Biomarkers important in the context of male fertility can be assessed using proton NMR What is known already Currently, the quality of sperm is assessed predominantly by examining its concentration, motility, and morphology. These parameters are insufficient in diagnosing male infertility and predicting treatment outcomes with assisted reproductive technology (ART). Metabolomics techniques have been shown to be potentially useful in identifying biomarkers related to infertility. It has been proven, using proton NMR, that there are a number of substances in the semen plasma that correlate with various physiological states of patients and may potentially indicate impaired functioning of various metabolic pathways. However, the relationship between biomarkers and embryo development has not yet been investigated Study design, size, duration This was a prospective case-control study and consisted of 30 semen samples from 13 patients who underwent IVF treatment between January – March 2022 at INVICTA Fertility Centre, Poland. Sperm motility, concentration, morphology, and DNA fragmentation of the samples were assessed. Before sperm preparation for fertilization, semen was frozen to perform 1H NMR analyses, which were preceded by centrifugation in order to separate the plasma. NMR analysis results were compared to the embryological results. Participants/materials, setting, methods It was possible to identify pre-selected biomarkers in semen plasma through the use of reference spectra and to quantify their concentrations using the internal standard addition method. Their relative concentrations (separately and in pairs) were collated with semen parameters, embryo development data and pregnancy outcomes. A Machine Learning approach was employed to identify the most important variables, which were then used for further modelling with the use of LGBM models. Main results and the role of chance NMR allows for the identification and measurement of biomarkers important in the assessment of the fertility potential of semen. The presence of amino acids, organic acids, sugar residues and fatty acid derivatives was confirmed by applying this technique. It was found that the concentration of these biomarkers is an individual feature - probably dependent on the physiological state of the organism. The results indicate that the ratios of biomarker concentrations to each other are more important than their individual concentrations. Machine learning modelling showed a strong correlation of several compounds with the chance of obtaining a TQ blastocyst. The amount of taurine, glutamic acid and choline together with its derivatives seem to be the most important factors in blastocyst development. The results indicate that if the amount of glutamic acid significantly surpasses glycerophosphocholine, choline exceeds taurine, the amount of taurine is higher than that of citrate, or the amount of choline is significantly higher than its phosphorus analogue the likelihood of obtaining a TQ blastocyst is higher. This would suggest that several metabolic pathways as the Krebs cycle, the one-carbon cycle or choline metabolism were modified signalling pathological conditions which the standard assessment of semen parameters cannot detect. Limitations, reasons for caution The NMR technique allows a fast determination of the content of many biomarkers, but results are in the mmol/l range. The results should be confirmed in a larger group of patients with different semen disorders Wider implications of the findings NMR allows for the rapid creation of unique metabolic profiles of patients, which can ensure the creation of personalized treatments for male infertility and the selection of sperm with the highest chance of obtaining a TQ blastocyst Trial registration number The studies were approved by the Ethics Committee at the Gdansk Regional Medical Board (No KB-3/22).

Crystal structure of 5-(β-d-gluco-pyran-osyl-thio)-N-(4-methyl-phen-yl)-1,3,4-thia-diazol-2-amine.

In the structure of the title compound, C15H19N3O5S2, the bond lengths at the linking sulfur atom are significantly different [1.7473 (17) and 1.811 (2) Å], and the angle at the exocyclic nitro-gen atom is wide at 128.45 (18)°. The inter-planar angle between the tolyl and thia-diazole rings is 9.2 (1)°. The complex hydrogen-bonding pattern, involving five donors and five acceptors, can be broken down into a one-dimensional ribbon parallel to the b axis, involving hydrogen bonds of the sugar residues only, and a two-dimensional layer structure parallel to the ab plane, based on the N-H⋯O and O-H⋯N hydrogen bonds.

Early in vitro evidence indicates that deacetylated sialic acids modulate multi-drug resistance in colon and lung cancers via breast cancer resistance protein.

Cancers utilize sugar residues to engage in multidrug resistance. The underlying mechanism of action involving glycans, specifically the glycan sialic acid (Sia) and its various functional group alterations, has not been explored. ATP-binding cassette (ABC) transporter proteins, key proteins utilized by cancers to engage in multidrug resistant (MDR) pathways, contain Sias in their extracellular domains. The core structure of Sia can contain a variety of functional groups, including O-acetylation on the C6 tail. Modulating the expression of acetylated-Sias on Breast Cancer Resistance Protein (BCRP), a significant ABC transporter implicated in MDR, in lung and colon cancer cells directly impacted the ability of cancer cells to either retain or efflux chemotherapeutics. Via CRISPR-Cas-9 gene editing, acetylation was modulated by the removal of CAS1 Domain-containing protein (CASD1) and Sialate O-Acetyl esterase (SIAE) genes. Using western blot, immunofluorescence, gene expression, and drug sensitivity analysis, we confirmed that deacetylated Sias regulated a MDR pathway in colon and lung cancer in early in vitro models. When deacetylated Sias were expressed on BCRP, colon and lung cancer cells were able to export high levels of BCRP to the cell's surface, resulting in an increased BCRP efflux activity, reduced sensitivity to the anticancer drug Mitoxantrone, and high proliferation relative to control cells. These observations correlated with increased levels of cell survival proteins, BcL-2 and PARP1. Further studies also implicated the lysosomal pathway for the observed variation in BCRP levels among the cell variants. RNASeq data analysis of clinical samples revealed higher CASD1 expression as a favorable marker of survival in lung adenocarcinoma. Collectively, our findings indicate that deacetylated Sia is utilized by colon and lung cancers to engage in MDR via overexpression and efflux action of BCRP.

Dammarane-type saponins from Gynostemma pentaphyllum and their anti-aging activities via up-regulating mitochondria related proteins

The importance of mitochondria in regulation of aging has been extensively recognized and confirmed. Gynostemma pentaphyllum (Thunb.) Makino, a homology of medicine and food, has been widely utilized as dietary supplement. In this study, the transcriptome of normal cells (wild type mouse embryo fibroblasts) regulated by the 30% aqueous EtOH extract of G. pentaphyllum was firstly evaluated by RNA sequencing and the results revealed that the G. pentaphyllum could up-regulate the genes involved in oxidative phosphorylation (OXPHOS) and sirtuin (SIRT) signaling pathways, indicating its effect in promoting cell viability might be attributed to the role of improving mitochondrial functions. To further discover the bioactive compounds, sixteen undescribed dammarane-type saponins along with twenty-eight known analogues were isolated from the active extract of G. pentaphyllum. Their structures were elucidated by means of comprehensive analysis of NMR and HRMS spectroscopic data. All isolates were evaluated for the regulatory effects on SIRT3 and translocase of the outer membrane 20 (TOM20), and thirteen of them exhibited satisfactory agonist activities on both SIRT3 and TOM20 at 5 μM. Furthermore, the preliminary structure-activity relationships analysis demonstrated the additional hydroxymethyl and carbonyl groups or less sugar residues in saponins could contribute positively to the up-regulatory effect on SIRT3 and TOM20. These findings encouraged the potential roles of G. pentaphyllum and its bioactive saponins in the development of natural drugs for the treatment of aging-related diseases.

Tumor cell-intrinsic and tumor microenvironmental conditions co-determine signaling by the glycoimmune checkpoint receptor Siglec-7

Tumors create an immunosuppressive tumor microenvironment by altering protein expression, but also by changing their glycosylation status, like altered expression of sialoglycans. Sialoglycans are capped with sialic acid sugar residues and are recognized by Siglec immune receptors. Siglec-7 is an inhibitory immune receptor similar to PD-1, and is emerging as glycoimmune checkpoint exploited by cancer cells to evade the immune system. However, the exact cellular and molecular conditions required for Siglec-7-mediated immune cell inhibition remain largely unknown. Here, we report on the development of a chimeric Siglec-7 cell system that enables dissection of Siglec-7 signaling, rather than Siglec-7 binding. Antibody-induced clustering, sialic acid-containing polymers, and highly sialylated erythrocytes effectively induced Siglec-7 signaling, thereby validating functionality of this reporter system. Moreover, the system reveals tumor cell-dependent Siglec-7 signaling. Tumor-associated conditions important for Siglec-7 signaling were defined, such as Siglec-7 ligand expression levels, presence of the known Siglec-7 ligand CD43, and sialic acid availability for sialylation of glycans. Importantly, therapeutic targeting of the Siglec-7/sialic acid axis using a sialyltransferase inhibitor resulted in strong reduction of Siglec-7 signaling. In conclusion, using a newly established cellular tool, we defined a set of tumor-associated conditions that influence Siglec-7 signaling. Moreover, the system allows to assess the efficacy of novel cancer drugs interfering with the Siglec-7/sialic acid axis as immunotherapy to treat cancer.

Increasing the Nutritional Value of FISH Feed with the Addition of Eco Enzyme

Feed is one of the determinants of the success of cultivation activities. The nutritional and protein content of commercial feed has been determined, however, the protein content in the feed can still be increased. Eco enzyme is the result of fermentation from a mixture of sugar, organic fruit or vegetable residue, and water by utilizing probiotic activity which produces enzymes from organic waste. The purpose of this study was to analyze the increase in the nutritional value of fish feed with the addition of eco enzyme. Analysis of feed samples was carried out at the Laboratory of Nutrition and Animal Feed Science, Faculty of Agriculture, Lambung Mangkurat University, South Kalimantan. The treatment given was the addition of eco enzyme at different doses with the treatment of feed A (20 mL/kg feed), B feed (40 mL/kg feed), and C feed (60 mL/kg feed). The organic ingredients used to make Eco Enzyme are fruit peels sourced from mini markets and salad traders in the area around Banjarbaru City. The results of the chemical feed analysis test (proximate) which include: water content, ash content, protein content, fat content and fiber content. ased on the results of the analysis of feed samples with the addition of 60 mL/kg of feed eco enzyme, the value of the protein content can increase by 117.4% from 15% protein content to 32.71%. It contains a fat content value of 3.33% which is close to the SNI requirements for fish feed quality and reduces 69.6% of the fiber content of feed from 10% to 3.04%.%.