Search Results (211)

Pompe disease is a rare autosomal-recessive neuromuscular disorder caused by a deficiency of the lysosomal enzyme acid alpha-glucosidase (GAA), leading to the pathological accumulation of glycogen and impaired autophagy. Enzyme replacement therapy (ERT) with recombinant human alpha-glucosidase (rhGAA) has been available since 2006, but may lead to the formation of anti-drug antibodies (ADAs) against the recombinant human enzyme, which, in turn, may adversely affect the response to ERT. Knowledge of the antigenic determinants of rhGAA involved in interaction with ADAs may facilitate the development of strategies to attenuate the anti-drug immune response in patients. Here, we determined the rhGAA ADA epitopes in the plasma of Pompe disease patients using a series of affinity purifications combined with epitope extraction and label free quantitation LC-MS. Full article

Amorpha fruticosa L. (Fabaceae) originates from North America and has become an aggressive invasive plant in many parts of the world. It affects the local biodiversity in many negative ways. Our previous in vivo tests of purified extract of A. fruticosa pods for antihyperglycemic activity in streptozotocin-induced diabetic spontaneously hypertensive rats (SHRs) revealed that the oral administration of purified extract of A. fruticosa (100 mg/kg) for 35 days to SHRs caused significant decreases in the systolic pressure, blood glucose levels, and MDA quantity. The aim of this experimental study is to test the glycosidase inhibition of several extracts of A. fruticosa pods. Methods: GC-MS, NMR, and a glycosidase inhibition assay were performed. Results: The results demonstrate strong inhibition of yeast alpha- and almond beta-glucosidases, rat intestinal hexosaminidase, and bovine beta-glucuronidase, but not of some other glycosidases. The activity is probably due at least in part to the presence of iminosugars and iminosugar acids. We here report on further analysis and activity assessments of A. fruticosa pods and leaves collected in Bulgaria, and for the first time discover glycosidase inhibitors, pinitol, and hydroxylated pipecolic acids in the species and more complex iminosugar-like compounds that may all contribute to antidiabetic potential. Hydroxylated pipecolic acids are probable precursors of iminosugars and common in legumes containing them. Considerable chemical variation was observed over four pod collections. Conclusions: A. fruticosa pods and leaves were found to contain a number of compounds that could contribute to the potential antihyperglycemic activities including pinitol and a complex mixture of iminosugar-related compounds derived from pipecolic acids and prolines. The pods and leaves caused potent selective inhibition of glucosidases and hexosaminidases and beta-glucuronidase. The variation between the collections might reflect the sites differing or wide phenotypic versatility allowing the success of the species as an invasive plant. Full article

Background: The New Zealand pine bark has been demonstrated in vitro to inhibit digestive enzymes involved in carbohydrate digestion (alpha-amylase, alpha-glucosidase, and dipeptidyl-peptidase 4 (DPP-4)). Objective: This study aims to investigate the inhibitory effects of the New Zealand pine bark on sucrose uptake and glycaemic responses in humans. Methods: A single-blind, randomised, placebo-controlled, crossover trial was carried out involving healthy adults (n = 40 (M: 12, F: 28), 30.1 ± 1.3 years, BMI 23.4 ± 0.5 kg/m², HbA1c 32.5 ± 0.6 mmol/mol, FBG 4.7 ± 0.1 mmol/L). A control (75 g of sucrose powder only), and two doses of the pine bark extract (50 and 400 mg) were provided on separate occasions, with 75 g of sucrose mixed in 250 mL of water. Blood samples were collected at −10, 0, 15, 30, 45, 60, 90, and 120 min via a finger prick test. A linear mixed model for repeated measures (SPSS v30, IBM) was applied, and data presented as model-adjusted mean ± SEM. Results: Compared to control (247.5 ± 14.0 mmol/L⋅min), the iAUCglucose was significantly reduced with the 400 mg dose (211.8 ± 13.9 mmol/L⋅min, 14.4% reduction, and p = 0.037), but not with 50 mg dose (220.8 ± 14.2 mmol/L⋅min, 10.8% reduction, and p = 0.184). Compared to control (9.1 ± 0.2 mmol/L), glucose peak value was significantly reduced with the 50 mg dose (8.6 ± 0.2 mmol/L, 5.5% reduction, and p = 0.016) but not with the 400 mg dose (8.7 ± 0.2 mmol/L, 4.4% reduction, and p = 0.093). There were no statistically significant changes in postprandial insulin levels with the pine bark extract compared to control. Conclusions: The New Zealand pine bark extract attenuated sucrose uptake with improved glycaemic responses, and may therefore be useful as a hypoglycaemic adjunct to the diet. Full article

This study evaluates the potential role of two extracts derived from green and roasted coffee in managing Type 2 Diabetes (T2D). The phytochemical analysis revealed that the roasted coffee extract (RCE) contains higher levels of flavonoids and tannins, while the green coffee extract (GCE) seems to be richer in phenolic acids. No differences in the antioxidant activity of both extracts were observed. The study demonstrated that GCE exhibits stronger prebiotic potential by significantly enhancing the growth of beneficial probiotic bacteria when compared to the untreated sample. Both extracts inhibited α-amylase and α-glucosidase enzymes, with RCE demonstrating a better performance than other commercial treatments for T2D. Glucose uptake assays on yeast cells demonstrated that both extracts enhance glucose transport, particularly at low glucose concentrations, reducing supernatant glucose levels by 60–80%. Notably, GCE maintained its effectiveness even at the highest glucose concentrations tested. These findings suggest that coffee extracts, particularly GCE, may be useful as nutraceuticals for potentially regulating glucose metabolism and gut microbiota in T2D management. Full article

Background and Aim: Rhoifolin is a bioactive flavonoid that possesses strong antioxidant and anti-inflammatory activities. The current investigation aimed to examine the anti-diabetic potential of rhoifolin in streptozotocin-induced diabetic rats. Dose-dependent (10 and 20 mg/kg) anti-hyperglycemic, anti-hyperlipidemic, anti-inflammatory, and antioxidant effects of rhoifolin were evaluated by measuring fasting blood glucose, serum glucose, serum insulin, HOMA-IR, lipidemic status, inflammatory cytokines, and hepatic antioxidant markers. To identify the underlying mechanism behind the anti-diabetic activity of rhoifolin, qRT-PCR was carried out using rat pancreatic and hepatic tissues. Results: The results have shown that rhoifolin produced antioxidant effects, as exhibited by DPPH and ABTS⁺ assays, respectively. Rhoifolin showed potent alpha-amylase and alpha-glucosidase inhibitory activities. Rhoifolin enhanced the serum insulin level, significantly decreased the serum glucose, HOMA-IR, and cytokine levels, and improved the lipid profile. Rhoifolin also showed a substantial decline in insulin resistance in the treated rats. Rhoifolin significantly raised catalase and superoxide dismutase levels in hepatic tissues while potentially decreasing the malondialdehyde levels. Moreover, rhoifolin significantly down-regulated the MAPK-8, TRAF-6, and TRAF-4 expressions and up-regulated the PDX-1, SIRT-1, INS-1, and GLUT-4 expressions in treated groups. Conclusions: Our results indicate that rhoifolin exhibits a hypoglycemic effect, which appears to be associated with its regulatory impact on metabolic inflammation and oxidative stress markers. This was accompanied by a lower HOMA-IR index, highlighting its potential role in promoting glucose homeostasis and mitigating insulin resistance. According to preliminary results, rhoifolin could further be tested to introduce it as another viable treatment option for diabetes. Full article

Pennsylvania started newborn screening for Pompe disease (PD) in 2016. As a result, the prevalence of PD has increased with early detection, primarily of late-onset Pompe disease (LOPD). No clear guidelines exist regarding if and when to initiate enzyme replacement therapy (ERT) in patients identified through a newborn screen (NBS). To help define the natural history and indications for starting ERT, we present the long-term follow-up data of 45 patients identified through NBS from 2016 to 2021. These patients were evaluated at regular intervals through our multi-disciplinary clinic at the Children’s Hospital of Philadelphia (CHOP) with physical examinations, physical therapy evaluations, muscle biomarkers including creatine kinase (CK), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and hexosaminidase 4 levels (Hex4), as well as cardiac evaluation at certain points in time. We found that newborn screening of acid alpha-glucosidase (GAA) enzyme detected primarily LOPD. One case of infantile-onset PD (IOPD) was detected. Muscle biomarkers in LOPD were elevated at birth and showed a general downward trend over time. NBS GAA levels and initial CK levels helped to differentiate LOPD cases from unaffected infants (carriers, pseudodeficiency alleles), while Hex4 was not a meaningful discriminator. On repeat NBS, there was a significant difference between mean GAA levels for the unaffected vs. compound heterozygote groups and unaffected vs. homozygote groups for the common splice site pathogenic variant (c.-32-13T>G). Echocardiogram and electrocardiogram (EKG) are essentially normal at the first evaluation in LOPD. One LOPD patient was started on ERT at age 4.5 months. Continued data collection on these patients is critical for developing management guidelines, including timing of ERT and improved genotype–phenotype correlation. Full article

Research on the effects of organic and inorganic Cu sources on metabolic processes and mechanisms in pigs is lacking. This study investigated the effects of different copper (Cu) sources and levels on hepatic Cu metabolism and transporter factors in growing pigs. Sixty healthy piglets (initial body weight 14.00 ± 0.30 kg) were randomly divided into four groups with five replicates of three pigs each. Four diets (AM, AH, BM, and BH) had different Cu sources [Cu sulphate (CuSO₄): A and Cu amino acids (Cu-AA): B] and levels [supplemented (120 mg/kg DM): M, supplemented (240 mg/kg DM): H]. The pre-feeding period was 7 days, followed by a 45-day feeding period. Slaughter and sample collection were carried out on the 46th day of the formal feeding period. Significant differences were considered at p < 0.05. The final weight and average daily gain (ADG) of growing pigs in the Cu-AA groups were significantly higher than those in the CuSO₄ groups. Serum Cu increased with increasing Cu supplementation on days 20 and 40. Cu concentrations in muscle, liver, and liver subcellular organelles were higher in Cu-AA groups. In the CuSO₄ groups, Cu concentrations were higher in kidneys and faeces. In Cu-AA groups, both the Cu concentrations in lysosomes and cytosol were higher, and the activities of cathepsin D (CTSD), β-glucosidase (BGL), and acid phosphatase (ACP) in lysosomes and cytoplasm were higher. Comparisons between groups showed that liver mRNA of copper transporter protein 1 (CTR1), ATPase copper-transporting beta (ATP7B), ceruloplasmin (CP), antioxidant protein 1 (ATOX1), and metallothionein (MT) was lower in the CuSO₄ group than in the Cu-AA group, with the best performance at 120 mg/kg Cu. mRNAs for ATPase copper-transporting alpha (ATP7A), cytochrome c oxidase copper chaperone 17 (COX17), and copper chaperone for superoxide dismutase (CCS) showed a decreasing trend in the Cu-AA groups. Cu-AA is better for Cu deposition, enhances the utilisation of Cu, reduces Cu excretion, and promotes the expression of relevant enzymes and transporters in the liver. Full article

(1) Background: As population growth accelerates, unsustainable practices such as excessive cutting and burning of desert plants in the transition zones between deserts and oases have led to widespread vegetation loss. (2) Methods: The experiment was conducted in the oasis transition zone on the southern edge of the Taklamakan Desert from 2010 to 2023 year. Among the treatments included a control group (CK), cutting in spring (CS), cutting in fall (CF), burning in spring (BS), and flood water irrigation (FI). We used high-throughput sequencing to determine soil microbial composition and diversity and routine laboratory methods to determine soil physical and chemical properties and enzyme activities. (3) Results: No significant differences in bacterial alpha diversity (Chao1, Dominance, Observed_features, Pielou_e, Shannon, and Simpson) across the different long-term disturbance patterns. In fungi, the CK treatment showed significantly higher Chao1, Shannon, and Observed_features indices compared to BS and FI. Principal component analysis revealed a substantial reduction in bacterial community diversity in BS compared to FI, while fungal communities were lower in CK and CS compared to BS, CF, and FI; (4) Conclusions: Soil moisture content, electrical conductivity, organic carbon, and the activity of the enzyme cellobiohydrolase as key factors shaping the bacterial community. For fungi, organic carbon and the β-1,4-glucosidase enzyme were the main drivers. Full article

Pompe disease is a neuromuscular disorder caused by a deficiency of the enzyme acid alpha-glucosidase (GAA), which leads to lysosomal glycogen accumulation and progressive development of muscle weakness. Two distinct isoforms have been identified. In the infantile form, the weakness is often severe and leads to motor difficulties from the first few months of life. In adult patients, the progression is slower but can still lead to significant loss of mobility. The current inherent difficulties of the disease lie in both early diagnosis and the use of biomarkers. Given that this is a multifactorial disease, a number of components may exert an influence on the disease process; from the degree of pre-ERT (enzyme replacement therapy) muscle damage to the damaged autophagic system and the different pathways involved. What methodology should be employed to study the complex characteristics of Pompe disease? Our approach relies on the application of genetic and epigenetic knowledge, with a progression from proteomics to transcriptomics. It is also becoming increasingly evident that artificial intelligence is a significant area of interest. The objective of this study is to conduct a comprehensive review of the existing literature on the known data and complications associated with the disease in patients with disorders attributed to Pompe disease. Full article

The present study examined the effect of the three different altitudes on the enzymatic activity and the prokaryotic communities of the rhizosphere of Petromarula pinnata (L.) A.DC. (Campanulaceae), a vulnerable local endemic species of Crete (Greece). It was observed that the pH and N-acetylglucosaminidase (NAG) activity increased with altitude while the β-1,4-glucosidase (BG) activity fluctuated with increasing altitude. The prokaryotic community in the rhizosphere of P. pinnata was dominated at the phylum level by Proteobacteria, Actinobacteriota, Bacteroidota, and Firmicutes, as well as by Bacillus members at the genus level. The alpha diversity did not vary with altitude while the b-diversity varied significantly, reflecting differences in community composition in relation to altitudinal gradient. The NAG activity was positively associated with most of the predominant phyla, except for Proteobacteria. The BG enzyme activity appeared to be negatively associated with Proteobacteria, Chloroflexi, and Acidobacteriota. Based on online databases, the predicted functions of the community showed a clear distinction in relation to altitude. At lower altitude, functions related to quorum sensing among microbes were overrepresented, while at the higher altitude, the functions were more related to energy production and transfer. The results of this research contribute to the ex situ and in situ protection of the vulnerable populations of P. pinnata and provide information for understanding the effect of altitude on processes in the rhizosphere of a threatened local endemic species of Crete studied in its original habitats. Full article

Green algae, particularly Ulva species, are rich in complex polysaccharides, such as ulvan, which have significant potential for biotechnological applications. However, the biochemical properties of ulvan depolymerised products remain underexplored. The enzymatic depolymerisation of ulvan has garnered attention owing to its cost advantages over alternative methods. Nevertheless, the biochemical characterisation of ulvan lyases, specifically those belonging to the polysaccharide lyase family 25 (PL25), is limited. In this study, we identified and biochemically characterised a novel PL25 ulvan lyase, PaUL25, which functions optimally at pH 10. Additionally, we explored the alpha (α)-glucosidase inhibitory properties of ulvan depolymerised products. PaUL25 exhibited optimum activity at 35 °C in Tris-HCl buffer (pH 10). Moreover, enzyme activity was enhanced by more than 150% in the presence of Mn²⁺ metal ions at and below concentrations of 10 mM. The endolytic action of PaUL25 produced ulvan oligosaccharides with degrees of polymerisation of 2 and 4 as its end products. Partially and completely hydrolysed ulvan oligosaccharides exhibited α-glucosidase inhibitory activity, with half inhibitory concentration IC₅₀ values of 3.21 ± 0.13 and 2.51 ± 0.19 mg/mL, respectively. These findings expand our understanding of PL25 and highlight the pharmaceutical potential of ulvan oligosaccharides, particularly as antidiabetic agents. Full article

The brown alga Sargassum fusiforme (SF) is historically consumed as a food material in Japan. A steaming process is often required for SF products on the market due to their moderate hardness and astringent taste. This investigation aimed to elucidate the effect of steaming on the anti-diabetic activity of SF and its related chemical components. Acetone extracts of SF were prepared after it were steamed for 0, 1, 2, or 4 h (SF-0h, SF-1h, SF-3h, and SF-4h, respectively). Alpha-glucosidase inhibitory profiles of each SF extract were made based on activity-guided separation. The active fractions were collected and NMR was applied for a further chemical composition analysis. Our results suggested that total polyphenol levels decreased drastically after steaming, which resulted in a drop in α-glucosidase inhibitory activity. The fatty acid, pheophytin a, and pyropheophytin a contents were elevated significantly after steaming, which contributed to the majority of the activity of steamed SF (SF-1h). However, prolonging the steaming time did not significantly affect the activity of SF further since the content of free fatty acids in steamed SF (SF-2h and SF-4h) almost did not change with a longer time of steaming. Moreover, palmitic acid, 8-octadecenoic acid, and tetradecanoic acid were identified as the top three important fatty acids for the inhibition of α-glucosidase by steamed SF. Further molecular docking results revealed that these fatty acids could interact with residues of α-glucosidase via hydrogen bonds, salt bridges, and hydrophobic interactions. In conclusion, steaming altered the α-glucosidase inhibitory properties of SF by changing the contents of polyphenols, fatty acids, and chlorophyll derivatives. Full article

Coffee silverskin (CS), the main by-product of coffee roasting production, contains various valuable bioactive compounds in its chemical compositions. Hydrothermal water extraction (HDTE) is one of the promising techniques for valorizing the organic fraction of CS into functional bioactive ingredients, which can be further exploited in various applications. This study aimed to evaluate the hyaluronidase and α-glucosidase inhibition activities of the CS extracts obtained under optimized water extraction conditions. Process optimization was performed using central composite design response surface methodology (CCD-RSM) with a broader range of extraction temperatures (25, 137.5, and 250 °C), reaction times (5, 38.5, and 72 min), and solid-to-liquid ratios (1:10, 1:80, and 1:150). The highest yield of 39.62% was obtained at 137.5 °C, with a reaction time of 72 min and an S/L ratio of 1:80. The total caffeoylquinic acid contents (T-CQA) were quantified based on the sum of three major isomers, including 3-CQA, 4-CQA, and 5-CQA. The results revealed that the highest T-CQA (2.76 ± 0.20 mg/g CS) was significantly obtained (p < 0.05) by subcritical water extraction (SWE) at 143.2 °C with an S/L ratio of 1:10 and an extraction time of 10.41 min. At such conditions, the total phenolic content (TPC), antioxidant properties (AP), and caffeine were 96.13 mg gallic acid equivalence per gram (GAE/g) CS, 20.85 ± 0.17 mg Trolox equivalence per gram (TE/g) CS, and 10.84 ± 1.25 mg/g CS, respectively. The 50% inhibition capacity (IC50) of hyaluronidase and α-glucosidase inhibition of the CS extracted were 5.00 mg/mL and 9.00 mg/mL, respectively. Our results supported the potential direct or indirect applications of CS, such as hydrothermal CS extract (HDT-CSE), in functional food or drinks. Repurposing CS residue to manufacture new products can efficiently reduce the amount of organic waste in landfills, thus conserving resources and energy and contributing to a lower overall carbon footprint in coffee production. Full article

Here, super porous carboxymethyl cellulose (CMC) cryogels were synthesized in 10–100% crosslinker and the presence of TA, at varying amounts of TA, e.g., 10 and 25 wt% of CMC under cryogenic conditions (−20 °C) as TA@CMCs. To control the degradation of CMC cryogel networks, the crosslinking ratio of divinyl sulfone (DVS:X) to CMC varied at 10, 25, 50, and 100% moles of the CMC repeating unit. Higher hydrolytic degradation was observed for CMC 10%X cryogels at pH 1.0 with 28.4 ± 1.2% weight loss. On the other hand, the TA-release studies from TA@CMC-based cryogels showed that higher TA releases were observed for both TA@CMC 10% and 25% cryogels at pH 7.4, with 23.6 ± 1.1, and 46.5 ± 2.3 mg/g in 480 min, which are equal to almost 24% and 18% of the TA contents of the corresponding cryogels, respectively. The antioxidant properties of TA@CMC cryogels were examined, and worthy antioxidant properties were observed due to the TA. The alpha-glucosidase enzyme inhibition ability of the prepared cryogels was examined at different concentrations by grinding cryogels, and it was determined that TA@CMC 25% cryogel at 3 mg/mL concentration inhibited 70.4 + 1.3% of the enzyme. All bare CMC-based cryogels were found to be non-hemolytic with a less than 1% hemolysis ratio and also effective on the blood coagulation mechanism with blood-clotting index (BCI) values between 62.1 and 81.7% at 1 mg/mL concentrations. On the other hand, TA@CMC 25% cryogels exhibited a slight hemolytic profile with a 6.1 ± 0.8% hemolysis ratio and did not affect the blood coagulation mechanism with 97.8 ± 0.4% BCI value. Full article

On commercial banana (Musa spp.) plantations, soils are often supplemented with phosphorus (P) fertiliser to optimise production. Such additions may influence the diversity and function of soil microbial communities, which play important roles in P cycling and affect plant fitness. Here, we characterised the effects of P addition on the diversity and function of banana-associated microbial communities. P addition was associated with significant increases in soil P and the activities of alpha-glucosidase, chitinase, arylsulphatase, and acid phosphatase, but not beta-glucosidase or xylosidase. P addition also expedited bunch emergence and harvest, but did not influence fruit yield, plant height, or foliar P. There were no significant effects of P addition on the alpha or beta diversity of bacterial, fungal, and nematode communities, including members of the core microbiome. The only exceptions to this was an increase in the relative abundance of a Fusarium population in roots. These results indicate that phosphorus application to banana soils may stimulate microbial enzyme activities with minor or negligible effects on microbial diversity. Full article