Introduction: Penicillin G amidase is an industrially significant enzyme widely employed in the production of semi-synthetic β-lactam antibiotics through the hydrolysis of Penicillin G to 6- aminopenicillanic acid. Owing to its commercial importance, extensive research has focused on improving the operational stability, reusability, and catalytic efficiency of PGA through various immobilization strategies. Methods: Optimization of multiple parameters for free and immobilized Penicillin G Acylase (PGA) is critical for improving the enzyme';s catalytic effectiveness, stability, and reusability in industrial and medicinal applications. This procedure entails methodically altering and analyzing variables such as substrate concentration, mechanical stability, cycle number, and storage conditions, and their effects on operational stability, pH, and temperature. PGA was optimized by entrapment on collagen hydrogel beads, resulting in collagen hydrogel + gelatin hybrid gel beads. Result: Immobilized PGA in Collagen Hydrogel + gelatin hybrid beads showed superior thermal stability, reusability, and storage stability as compared to gelatin-immobilized PGA. The entrapment of PGA onto Collagen Hydrogel + gelatin hybrid beads revealed several advantages and could be used in the production of 6-aminopenicillanic acid (6APA). Discussion: The study investigated the biochemical behavior of Penicillin G amidase (PGA) immobilized on collagen hydrogel and a collagen–gelatin bio-composite. Relative analysis focused on enzyme activity, stability, and mechanical strength, revealing insights into their appropriateness as immobilization matrices for enhanced PGA performance in industrial biocatalysis applications. Conclusion: Hydrogel + gelatin hybrid beads are more beneficial in industrial applications due to their greater stability and usability. PGA entrapment onto Hydrogel + gelatin hybrid beads has shown numerous advantages and may be useful in the manufacture of 6APA (6-aminopenicillanic acid).

Abstract: Metabolic disruption associated with obesity leads to a persistent, subclinical inflammatory state called metabolic inflammation, which in turn further exacerbates dysmetabolism. Notably, during obesity, there is also a remarkable hypoxia. The prevailing understanding is that hypoxia in obesity is primarily caused by decreased cardiac output and blood flow, as well as an increase in adipocyte diameter. Consequently, hypoxia contributes to metabolic inflammation by regulating metabolic and inflammatory pathways. However, in this perspective, we propose another mode of obesity- induced hypoxia. Aberrant systemic metabolism affects erythrocyte metabolism, impairing the production of 2,3-bisphosphoglycerate (2,3 BPG), the primary negative allosteric regulator of hemoglobin. This would lead to tissue hypoxia due to decreased oxygen release. It has been found that during obesity, elevated glucose is diverted towards sorbitol production and the synthesis of advanced glycation end products (AGEs). This diversion surpasses the increased glucose utilization by the pentose phosphate pathway, leading to the formation of reactive oxygen species. In turn, the oxidized Band 3 protein limits the rate of glycolysis, thereby reducing the synthesis of 2,3-BPG. Concurrently, reduced ATP levels impair de novo glutathione synthesis, despite an increased availability of amino acids, thereby exacerbating oxidative stress. ROS can also activate arginase, which in turn promotes further ROS generation by uncoupling nitric oxide synthase. Ultimately, increased systemic lipid accumulation leads to increased ceramide content, thereby inhibiting the adenosine monophosphate- dependent kinase, which regulates the activity of BPG mutase. Altogether, these mechanisms would hinder oxygen release by erythrocytes, leading to hypoxia.

Introduction: Diabetes mellitus is a rapidly escalating global health concern, and numerous ethnobotanical remedies are under investigation for their antidiabetic properties. Tinospora sinensis has long been used in traditional medicine by indigenous populations for glycemic control. This study aimed to characterize the phytochemical profile of T. sinensis and evaluate its in vitro α- glucosidase inhibitory activity, as well as assess its antioxidant and antimicrobial activities. Methods: Standard qualitative assays were used to identify secondary metabolites. Total phenolic content (TPC) was quantified via Folin–Ciocalteu reagent (expressed as mg gallic acid equivalents [GAE]/g), and total flavonoid content (TFC) via aluminum chloride colorimetry (mg quercetin equivalents [QE]/g). Antioxidant potential was determined using the DPPH radical scavenging assay. α-Glucosidase inhibition was measured spectrophotometrically using p-nitrophenyl-α-Dglucopyranoside as a substrate. Antimicrobial efficacy against Staphylococcus aureus was tested using the agar well diffusion technique. Results: Phytochemical screening confirmed the presence of alkaloids, flavonoids, glycosides, terpenoids, saponins, phenols, tannins, steroids, and quinones. TPC and TFC values were 181.41 ± 1.57 mg GAE/g and 12.08 ± 0.11 mg QE/g, respectively. The methanolic extract demonstrated considerable antioxidant activity (DPPH IC₅₀ = 111.43 ± 1.13 μg/mL). Discussion: Both crude and ethyl acetate extracts exhibited significant α-glucosidase inhibition (comparable or superior to control). Notable antibacterial activity was observed against S. aureus, with a 9 mm inhibition zone. result: The qualitative phytonutrient screening of T. sinensis reveals the presence of various biochemical compounds, including alkaloids, flavonoids, glycosides, terpenoids, saponins, phenolic groups, tannins, steroids, and quinones. The total phenolic content (TPC) and total flavonoid content (TFC) in the crude extract of T. sinensis were determined to be 181.41 ± 1.57 mg GAE/g and 12.08 ± 0.11 mg QE/g, respectively. Among the extracts from three different fractions, the methanolic extract demonstrated the strongest antioxidant activity with an IC50 value of 111.43 ± 1.13 µg/mL. The crude extract of T. sinensis exhibited significant α-glucosidase inhibitory activity with an IC50 value of 36.40 ± 0.35 µg/mL. Similarly, the ethyl acetate fraction from T. sinensis showed α-glucosidase inhibition with an IC50 value of 34.02 ± 1.13 µg/mL. Additionally, this plant displayed antimicrobial activity against Gram-positive Staphylococcus aureus, with a zone of inhibition (ZOI) measuring 9 mm. Conclusion: The presence of diverse bioactive phytochemicals in T. sinensis supports its traditional use in diabetes management. Its potent α-glucosidase inhibition suggests a mechanism for attenuating postprandial hyperglycemia, while antioxidant and antimicrobial activities substantiate additional therapeutic roles. These findings warrant further in vivo studies and mechanistic exploration to validate its potential as a source of enzyme inhibitors and therapeutic agents for metabolic disorders.

Introduction: The present study aimed at studying the potential of methanolic extract of Muntingia calabura bark (MBE) to inhibit acetylcholinesterase (AChE) in vitro Methods: Acetylcholinesterase (AChE) activity was assessed using chicken brain homogenate as the enzyme source. The assay was performed using acetylthiocholine iodide as a chromogenic substrate, and the reaction was monitored kinetically at 420 nm by measuring the rate of substrate hydrolysis. Results: MBE was found to inhibit the AChE activity with an IC50 value of 78.6±2.3 µg/mL. Analysis of the double reciprocal Lineweaver-Burk plot revealed that the rate of substrate hydrolysis by the brain homogenate was characterized by the Km and Vmax values of 93.7±18.8µM and 0.145±0.009 (delta OD/min at 412nm), respectively. In the presence of the MBE, we observed Km and Vmax values of 76.5±8.9 (without statistical difference compared to the control) and 0.07±0.007 (delta OD/min at 412 nm, statistically lower than the control), respectively, indicating that the MBE non-competitively inhibits AChE. Discussion: The data presented herein suggest that MBE inhibits AChE in vitro. Additional experiments are required to establish oral availability, toxicity, and efficacy in vivo. Conclusion: Our work demonstrates the potential of MBE to inhibit AChE in vitro and suggests that the extract warrants further exploration for molecular characterization and potential usefulness in mitigating pathologies in animal models of Alzheimer's disease.

Abstract: Type 2 diabetes mellitus is a growing global public health issue, with its prevalence projected to increase in the coming decades. It is one of the most prevalent and growing global health concerns, affecting millions of individuals worldwide. The condition is classified into two primary types: Type 1 diabetes, an autoimmune disorder that leads to the destruction of insulin-producing beta cells in the pancreas, and Type 2 diabetes, which is predominantly associated with insulin resistance and inadequate insulin secretion. The various enzymes play a crucial role in the regulation of metabolic pathways, and their dysfunction can contribute to various diseases, including diabetes mellitus. Among these enzymes, the dipeptidyl peptidase-4 serves as a therapeutic target for managing T2D. Inhibiting DPP-4 prevents the breakdown of glucose-dependent insulinotropic peptide and glucagon-like peptide 1, thereby maintaining their natural levels and helping to reduce blood glucose. This review provides a comprehensive overview of the DPP-4 enzyme, including the effects of DPP-4 inhibition on pancreatic beta cell function, skeletal muscle function, and glucose-lowering mechanisms. We believe that this information will aid scientists in developing novel antidiabetic compounds for T2D treatment.

Abstract: Sirtuin 1 (SIRT1), an NAD+-dependent deacetylase, plays a vital role in neurodegenerative diseases, particularly Alzheimer’s disease (AD). SIRT1 exerts neuroprotective effects by modulating oxidative stress, neuroinflammation, and mitochondrial function while promoting neuronal survival. It enhances amyloid-β (Aβ) clearance by activating α-secretase and inhibiting β-secretase, thereby reducing Aβ aggregation. Beyond protein aggregation, SIRT1 also influences mitochondrial biogenesis and function via peroxisome proliferator-activated receptor-gamma coactivator-1 alpha (PGC-1α), thereby enhancing energy metabolism and reducing oxidative damage. Moreover, SIRT1-mediated deacetylation of nuclear factor kappa B (NF-κB) suppresses neuroinflammation, further contributing to neuroprotection. Preclinical studies highlight the therapeutic potential of SIRT1 activators, such as resveratrol, in improving cognitive function and reducing AD pathology. However, challenges such as bioavailability, pharmacokinetics, and the lack of definitive clinical validation hinder its therapeutic translation. Emerging research suggests that lifestyle factors, including caloric restriction, exercise, and sleep modulation, may naturally enhance SIRT1 activity, offering alternative approaches for AD prevention. Despite promising evidence, further research is required to fully elucidate the mechanisms underlying SIRT1’s role in AD and to develop effective, targeted therapies. This review underscores SIRT1’s multifaceted involvement in AD pathophysiology and its potential as a therapeutic target, calling for multidisciplinary efforts to advance its clinical application.

Aims: This study aimed to determine the anti-diabetic potency of Hygrophila auriculata. Additionally, it assessed the in silico pharmacokinetic properties, drug-likeness, and toxicity profiles using SwissADME, ProTox II, ADMETlab2.0, and pkCSM web tools. Background: Diabetes mellitus (DM) is rising alarmingly, necessitating effective natural therapies to complement conventional treatments. Traditionally used in Ayurveda, Hygrophila auriculata exhibits multifaceted benefits, including hepatoprotection, antibacterial, antitumor and anti-diabetic properties, making it a promising natural remedy for diabetes. Methods: Dried and powdered Hygrophila auriculata underwent extraction and resulting methanolic fraction was utilized for analysis of total phenol and flavonoid content, 2,2-Diphenyl-1- picrylhydrazyl (DPPH) and superoxide anion scavenging assays, α-amylase and α-glucosidase enzyme inhibition assays. Molecular docking studies were also carried out. The drug-likeliness of the phytoconstituents present in H. auriculata was examined and ADMET properties were predicted using various web servers. Results: Hygrophila auriculata exhibited significant phenol (10.11±0.0025 mg GAE/g sample) and flavonoid content (102.05±0.0053 mg QE/g sample) and potent antioxidant activity, evidenced by DPPH radical scavenging assay (IC50 = 61.69 ± 0.133 µg/mL) and superoxide anion scavenging assay (IC50 = 34.91 ± 0.115 µg/mL). The extract also showed enhanced α-amylase inhibition (IC50 = 35.72±0.121 µg/mL) and robust α-glucosidase inhibition (IC50 = 64.42 ± 0.107 µg/mL). Using the phytocompounds, a thorough molecular docking study against target proteins linked to diabetes mellitus revealed that apigenin-7-O-glucuronide and cosmosiin were the most effective phytochemicals. Conclusion: The results suggest that Hygrophila auriculata possesses significant anti-diabetic properties, supporting its potential as a promising future anti-diabetic drug.

Abstract: The text discusses the critical role of enzyme immobilization in enhancing the efficiency, reusability, and stability of biocatalysts in industrial applications. Immobilization techniques include covalent bonding, encapsulation, adsorption, and cross-linking, each with its unique advantages and challenges. Covalent bonding ensures strong, irreversible attachment of enzymes to supports, preventing leaching and maintaining enzyme stability under various conditions. Encapsulation protects enzymes within a semi-permeable matrix, preserving their activity while allowing access to substrates. Adsorption, relying on weak interactions, is simple and reversible but prone to enzyme leaching. Cross-linking involves intermolecular bonding between enzymes and supports, enhancing stability but potentially altering enzyme conformation. Selecting appropriate supports—organic or inorganic—is crucial to minimize enzyme deactivation and maintain activity. Organic supports, like chitosan and alginate, offer biocompatibility and sustainability, while inorganic supports, such as silica and metal oxides, provide robustness and high surface areas. The text highlights the significance of optimizing immobilization techniques for specific enzymes, considering factors like mechanical resistance, substrate diffusion, and compatibility with enzyme structures. Recent advancements include the development of novel supports like hybrid materials and the application of nanotechnology, which offers enhanced stability and catalytic properties. However, challenges like enzyme deactivation, activity loss over time, and high immobilization costs persist. The text emphasizes ongoing research to address these issues, aiming to improve the economic viability and efficiency of immobilized enzymes in industrial processes. The study underscores the importance of tailoring immobilization strategies to specific enzymes and applications, ensuring maximal catalytic performance and reusability.

Abstract: Cancer is characterized by the uncontrolled proliferation of abnormal cells that escape the body's standard regulatory mechanisms. Under normal conditions, cells grow, divide, and die in an orderly manner, but cancerous cells lose this control, growing uncontrollably and invading surrounding tissues. Poly(ADP-ribose) polymerase 1 (PARP1) is a crucial enzyme in this DNA repair process, helping to fix single-strand breaks. PARP inhibitors (PARPi) are a class of drugs that target and block the activity of the PARP1 enzyme, impairing its ability to repair DNA damage. By inhibiting PARP1, these drugs lead to an accumulation of DNA damage in cancer cells, which eventually becomes overwhelming and leads to cell death. This mechanism is particularly effective in cancers with deficiencies in other DNA repair pathways, such as those with BRCA1 and BRCA2 gene mutations. Several PARPi, including Olaparib, Niraparib, and Rucaparib, have been approved by the U.S. Food and Drug Administration (FDA) for use in treating cancers like breast, ovarian, and prostate cancer, particularly in patients with BRCA mutations. The evolution and development of PARP inhibitors have focused on modifying their chemical structure to increase their effectiveness. The design of PARPi also aims to improve their bioavailability, ensuring that the drugs are effectively absorbed into the body and can reach the tumor site in sufficient concentrations. Further developments may also involve combining PARPi with other treatments, such as chemotherapy or immunotherapy, to enhance their overall efficacy.

Introduction: Liposomes are versatile drug delivery vehicles due to their nanoscale lipid bilayer vesicles, capable of encapsulating both hydrophilic and hydrophobic substances. They have shown promise in vaccine development, gene therapy, cancer treatment, and targeted drug delivery. However, their clinical applicability is limited due to factors like drug stability, manufacturing constraints, regulatory challenges, and immune responses. This study explores liposome formulations by focusing on enhanced stability, robustness, and drug-loading efficiency. It also discusses therapeutic implementation challenges. Methods: A systematic literature review was conducted using specific keywords and Boolean operators across databases, such as Web of Science, PubMed, and Scopus. Non-peer-reviewed articles, conference abstracts, and studies with poor methodology were excluded. Results: This review highlights advances in liposome formulation that boost therapeutic performance, enhance stability, and improve drug loading. Despite their promise, clinical application depends on overcoming issues like manufacturing complexity, regulatory constraints, and immune reaction limitations. Discussion: Liposomes enable efficient encapsulation and targeted delivery for both hydrophilic and hydrophobic drugs, enhancing therapeutic efficacy. Their biocompatibility makes them effective in cancer therapy, vaccine transport, and gene delivery. Nevertheless, further research is needed to improve production processes and ensure long-term safety for regulatory approval and commercial scalability. Conclusion: Liposomes hold strong potential for medical use and drug delivery. To achieve broader clinical adoption, challenges in formulation and regulation must be addressed. This review highlights recent innovations and strategies to optimize liposome-based therapeutics.