Physicochemical Assays Research Articles

Closing the Design-Make-Test-Analyze Loop: Interplay between Experiments and Predictions Drives PROTACs Bioavailability.

The drug development landscape is expanding to include drug modalities such as PROteolysis-TArgeting Chimeras (PROTACs) and peptides, offering possibilities for previously intractable biological targets. However, with their size and chemical nature, they diverge from established frameworks for the prediction of oral bioavailability. This evolution to larger and more complex molecules necessitates new methodologies and prediction models to continuously expand on bioavailability guidelines. We describe the high-capacity adoption of two chromatographic physicochemical assays and their application for iterative compound optimization to achieve oral bioavailability. We further describe how these data underpin the continuous refinement of internal machine learning models, which guide compound synthesis decisions in the molecular design phase. Based on data for a set of 691 PROTACs, and two project examples, we confirm a sweet spot for oral bioavailability at log D values higher than the norm for small molecules and show how experimental data and prediction models synergize to effectively drive chemistry optimization.

Indigenous Lactic Acid Bacteria as Antioxidant Agents in the Production of Organic Raw Fermented Sausages

The study aimed to assess the impact of lactic acid bacteria (LAB) strains on the antioxidant, physico-chemical properties, and microbiological quality of fermented sausages. Five treatments of raw sausages were prepared: two controls without LAB addition (C, P), and three samples with LAB addition (SCH1, BAL6, KL14). Fatty acid composition, cholesterol content, physico-chemical, microbiological tests, and antioxidant assays, were performed at time 0 and after 1 and 2 months of storage. A significantly higher ability to scavenge free radicals of DPPH (2,2-diphenyl-1-picrylhydrazyl) was found in sausages with all LAB strains. In the case of the ABTS (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) test, it was noted that KL14 treatment had higher antioxidant activity. The main fatty acids in sausages were monounsaturated and saturated. A significantly lower cholesterol content was observed in sausages with the addition of LAB. Sausages with LAB strains differed significantly in pH value. Water activity decreased significantly during storage. After 2 months of storage, the sausages with BAL6 and KL14 strains were characterized by significantly lower redox potential and a lower TBARS (thiobarbituric acid reactive substances) index. It was found that P sausages had the darkest color. SCH1, BAL6, and KL14 strains were also capable of producing red color. The total number of microorganisms in the sausages was high, which is mainly due to the high LAB content and yeast and mold counts. No spoilage or pathogenic microflora were detected. Indigenous LAB strains have the potential to improve the quality and safety of fermented meat products.

Exploring the Impact of In Vitro-Transcribed mRNA Impurities on Cellular Responses.

Advances in mRNA technology have enabled mRNA-based therapies to enter a new era of medicine. Such therapies benefit from a single, standardized in vitro transcription (IVT) manufacturing process applicable to a wide range of targets. This process includes several downstream purification steps, which aim to eliminate impurities that potentially affect safety and efficacy. However, it is not fully understood which impurities are the most critical; hence, some efforts are still needed to establish the correlation between RNA impurities and their role in limiting therapeutic efficacy. To study this relationship, we produced in vitro-transcribed mRNAs using several bacteriophage T7 RNA polymerases, including one wild-type and four engineered variants. Important attributes of the mRNA such as integrity, purity, and functional activity were then measured using advanced physicochemical and cellular assays. For impurities including abortive transcripts, mRNAs containing partial poly(A) tails, and double-stranded (ds)RNA byproducts, structure-function relationships have been established by tracking cellular responses (i.e., protein expression, reactogenicity) in multiple cell models. By varying the T7 RNA polymerase, different levels of sense-antisense dsRNA byproducts were measured by mass photometry, contributing directly to immunological reactogenicity in bone marrow-derived dendritic cells. T7 RNA polymerase differences with regard to short (<20 nucleotides) 3'-loopback dsRNA byproducts were also further investigated using native mass spectrometry by precisely resolving these impurities at the nucleotide level. Overall, this study highlights the importance of developing sensitive and advanced analytical methods to characterize IVT mRNA impurities and understand their interaction with cellular machinery in order to ensure quality control of RNA-based therapies.

A potent antimicrobial glycolipopeptide GLIP and its promising combined antimicrobial effect

Here, the glycolipopeptide GLIP was obtained by coupling IL-C8 and the monosaccharide molecule D-(+)-glucosamine to the N-terminal and C-terminal of the peptide P, which was designed on the basis of the biological characteristics of the antimicrobial peptides. In vitro bioactivity and physicochemical properties assays confirmed that GLIP had excellent antimicrobial activity against Gram-negative E. coli ATCC 25922 and Gram-positive S. aureus ATCC 29213, as well as good stability in serum and trypsin, low hemolysis, and good bacterial membrane-disrupting ability. In addition, the glycolipopeptide GLIP could self-assembly in aqueous solution to form spherical nano-aggregates, which could encapsulate the small molecule antibiotic TC to form the nanomedicine GLIP@TC and release the TC continuously and slowly in a sustained-release manner, exerting the combined antimicrobial effect of both. The results of animal experiments demonstrated the excellent in vivo antimicrobial activities of GLIP and nanomedicine GLIP@TC. Finally, molecular docking experiment showed that the GLIP could effectively bind to penicillin-binding protein 5 (PBP5) of E. coli and possibly inhibit its D-Ala carboxypeptidase (CPase) activity. All these results may imply the great potential of GLIP for clinical application against bacterial drug resistance.

Site-specific sulfations regulate the physicochemical properties of papillomavirus-heparan sulfate interactions for entry.

Certain human papillomaviruses (HPVs) are etiological agents for several anogenital and oropharyngeal cancers. During initial infection, HPV16, the most prevalent cancer-causing type, specifically interacts with heparan sulfates (HSs), not only enabling initial cell attachment but also triggering a crucial conformational change in viral capsids termed structural activation. It is unknown, whether these HPV16-HS interactions depend on HS sulfation patterns. Thus, we probed potential roles of HS sulfations using cell-based functional and physicochemical assays, including single-molecule force spectroscopy. Our results demonstrate that N-sulfation of HS is crucial for virus binding and structural activation by providing high-affinity sites, and that additional 6O-sulfation is required to mechanically stabilize the interaction, whereas 2O-sulfation and 3O-sulfation are mostly dispensable. Together, our findings identify the contribution of HS sulfation patterns to HPV16 binding and structural activation and reveal how distinct sulfation groups of HS synergize to facilitate HPV16 entry, which, in turn, likely influences the tropism of HPVs.

The potential effect of peel extracts of banana varieties: an in-vitro assessment

Background: Assamese cuisine is known for its use of kolakhar, a traditional ingredient; Rhizome, the skin and stem of bananas, can be used to make it. An ash filter from a banana tree is used to produce antacids. The word: kol” or “kola” is a local term for banana. In Assam, India, kolakhar is a common food additive and traditional ingredient. Method: Water is filtered through banana tree ashes to create this. The banana peel is burned after it has been dried. The ash is subsequently blended with water and left overnight. The mixture is filtered through a fine cloth once the ash has settled to the bottom of the container by the following morning. Several studies were carried out by evaluating the preparation of peel extract, Physicochemical parameters, antioxidant activity, etc. followed by some analytical methods to find the biologically active components, potential uses, and additional benefits of banana peels beyond what they currently serve as waste products. Finally, an antimicrobial study was performed by using the disc diffusion method. In this study, 4 different types of banana species investigated sought to determine the antioxidant capacity, antimicrobial activity, FT-IR, and UV to determine which one is better. Result: The physicochemical parameters, analytical technique, and assay provide an overview of the chemical characteristics, phytoconstituents, and food safety of kolakhar, which contribute to its unique properties in both traditional medicine and culinary applications. Conclusion: In conclusion, depending on the banana type used, banana peel extracts exhibit considerable promise as organic antioxidants and antibacterial agents. Therefore, considering all parameters, we obtained various potential effects from this study. The results are discussed with a graphical representation of the banana peel extract.

Evaluation of candidate International Standards for meningococcal capsular polysaccharide groups W and Y

Two candidate International Standards for meningococcal capsular group W and Y (MenW and MenY, respectively) polysaccharides were assessed for their suitability as quantitative standards in various physicochemical assays. The study was designed to evaluate the intended purpose of these standards, namely, to standardize the quantification of the respective polysaccharide content in meningococcal polysaccharide and conjugate vaccines and their intermediate components. Twelve laboratories from eleven different countries participated in the collaborative study of candidate preparations for International Standards for MenW and MenY polysaccharide (coded 16/152 and 16/206, respectively). Unitage was assigned using the Resorcinol assay. Our proposals, on the basis of data from the Resorcinol assay were: 1) candidate standard for MenW polysaccharide (16/152) to be assigned a content of 1.015 ± 0.071 mg MenW polysaccharide per ampoule (expanded uncertainty with coverage factor k = 2.13, corresponding to a 95 % level of confidence) and 2) candidate standard for MenY polysaccharide (16/206) be assigned a content of 0.958 ± 0.076 mg MenY polysaccharide per ampoule (expanded uncertainty with coverage factor k = 2.26, corresponding to a 95 % level of confidence). The amount of polysaccharide per ampoule remained consistent under all stability conditions over a 36-month period.

Preparation of arbutin hydrogel formulation as green skin lightener formulation: in vitro and in vivo evaluation

A hydrogel formulation containing arbutin was developed for use as a topical cosmetic product aimed at enhancing the anti-melanogenesis effect and skin delivery. To develop Arbutin-hydrogel, 1% arbutin was incorporated into a Chitosan/alginate hydrogel. The impacts of biopolymer proportion on the arbutin-hydrogel preparations were investigated. Swelling analysis, weight loss analysis, differential scanning calorimetry (DSC), scanning electron microscopy (SEM), attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy, toxicity study and, skin absorption test were employed to assess the arbutin-hydrogel. The swelling percentages of arbutin-hydrogel increased significantly after 4 h. In vitro, cytotoxic activity revealed that arbutin-hydrogel has no toxicity. Also, the cutaneous penetration investigation revealed that arbutin-hydrogel had a superior cutaneous accumulation (42.25 ± 0.30%) compared to plain gel (16.80 ± 0.33%). Also, physico-chemical specifications assay and solid state assessment were employed to assess of optimum formulation. Arbutin-hydrogel (60.9 ± 1.68%) inhibited melanin formation more effectively than arbutin solution. In addition, arbutin-hydrogel exhibited a more inhibitory impact on L-dopa auto-oxidation (54.90 ± 2.26%) than arbutin solution (38.62 ± 2.26%). In addition, the Wistar rat cutaneous sensitivity testing demonstrated that the hydrogel component did not affect the epidermis. These outcomes demonstrated that the arbutin-hydrogel could potentially be utilized for Arbutin topical delivery, expanding the treatment options for hyperpigmentation disorders.

Optimised Formulation of a New Sweet Apricot Kernel-Enriched Yoghurt: Assessment of Physicochemical, Sensory and Antioxidant Properties.

The addition of sweet apricot kernel powder, a by-product of apricot processing, to yoghurt appears to be particularly interesting option for the innovation of new food products. This study focuses on the formulation of a novel yoghurt enriched with sweet apricot kernel powder, sugar and milk powder. Different yoghurts were prepared by mixing sweet apricot kernel powder, sugar and milk powder as ingredients based on the simplex-centroid mixture design. The optimisation process took into account the physicochemical, antioxidant and sensory properties of the yoghurt. The results showed that the optimum values of sugar, milk powder and apricot kernel powder were 3.07, 2.16 and 2.77 %, respectively. The physicochemical assays showed that the addition of apricot kernel powder led to a significant increase in total phenolic content, antioxidant activity, syneresis, viscosity and acidity. The addition of sugar and milk powder also had a significant effect on the taste, texture and consistency of the yoghurt. Moreover, the enrichment of the product with apricot kernel powder significantly influenced the colour, odour, taste, texture and consistency. In conclusion, the optimised yoghurt enriched with apricot kernel had an interesting phenolic content and antioxidant properties with sensory acceptability, while reducing the amount of sugar and milk powder. This confirms the potential of using sweet apricot kernels as an ingredient in yoghurt production. The use of a simplex-centroid mixture design to optimise a new yoghurt formulation enriched with sweet apricot kernels shows significant improvements in total phenolic content, antioxidant activity and sensory acceptability. In addition, less sugar and milk powder is needed. The addition of sweet apricot kernels to yoghurt is therefore a new approach to improving its nutritional value and sensory appeal.

An Automated Robotic Interface for Assays: Facilitating Machine Learning in Drug Discovery by the Automation of Physicochemical Property Assays.

Measuring the physicochemical properties of molecules is an iterative but integral process in the drug development process. A strategy to overcome the challenges in maximizing assay throughput relies on the usage of in silico machine learning (ML) prediction models trained on experimental data. Consequently, the performance of these in silico models are dependent on the quality of the utilized experimental data. To improve the data quality, we have designed and implemented an automated robotic system to prepare and run physicochemical property assays (Automated Robotic Interface for Assays, ARIA) with an increase in sample throughput of 6 to10-fold. Through this process, we overcame major challenges and achieved consistent reproducible assay data compared to semiautomated assay preparation.

Product development and characterization of the Ayurvedic herbo-mineral-metallic compound- Hridayarnava Rasa

BackgroundHerbo-mineral-metallic formulations are an inseparable part of the Ayurveda system of traditional medicine. Hridayarnava Rasa (HR) is a preparation containing metals like copper, sulphur, and mercury in processed forms and other herbs that do not produce toxic effects and adverse drug reactions when taken in appropriate dosage. Ayurveda practitioners use it in treating cardiac diseases like hypertension, cardiotoxicity and many more. The rasa-aushadhis possess characteristics such as rapid efficacy, little dosage required, and extensive therapeutic applicability. Hridayarnava Rasa [AFI Part-1, 20:55] has been employed for the treatment of various diseases from ancient times. A systematic study of these formulations manufacturing is required to maintain their quality, safety, and efficacy is a need of time to protect the immense faith of patients in Ayurveda. ObjectivesThe present study aimed to prepare HR as per standard operating procedures mentioned in the classical text and to characterize it physio-chemically using advanced analytical techniques. Materials and MethodsHR was prepared and physicochemical analyses and assay of elements by ICP-AES were carried out as per Ayurvedic Pharmacopoeia of India (API). Powder X-ray diffraction (XRD), Field emission gun scanning electron microscopy with energy dispersive spectroscopy (FEG SEM, EDAX), CHNS-O analysis, Fourier Transform Infrared Spectroscopy (FTIR), Thermo-gravimetric analysis (TGA), Particle size distribution analysis (PSD) was carried out. ResultsThe XRD analysis of HR showed the presence of unreacted sulphur and sulfides of copper and mercury. FEG SEM revealed the particles in the form of aggregates as nanocrystallites in the range of 100–1000 nm. Elemental analysis showed the presence of copper, sulphur, and mercury in major, along with traces of iron, calcium, sodium, potassium, and magnesium. In FTIR analysis, 18 peaks were observed, which strongly suggests the presence of various organic groups. In the TGA, four peaks were seen, which can be attributed to sulphur volatilization and oxidative changes in mercury. In PSD analysis, 50% of the material was found below 16.40 μm. ConclusionTo establish a piece of fundamental knowledge and ensure uniformity of these rasa-aushadhis, it is imperative to conduct an analysis of their characteristics as per classical texts and modern analytical techniques. Additionally, it is crucial to investigate the significance of each procedural step included in the preparation process. The inferences drawn are helpful as an essential aid for quality assurance and standardization of this herbo-mineral-metallic formulation.

Hydrophobic solution functions as a multifaceted mosquito repellent by enhancing chemical transfer, altering object tracking, and forming aversive memory

Developing a safe and potent repellent of mosquitoes applicable to human skins is an effective measure against the spread of mosquito-borne diseases. Recently, we have identified that hydrophobic solutions such as low viscosity polydimethylsiloxane (L-PDMS) spread on a human skin prevent mosquitoes from staying on and biting it. This is likely due to the ability of L-PDMS in wetting mosquito legs and exerting a capillary force from which the mosquitoes attempt to escape. Here we show three additional functions of L-PDMS that can contribute to repel Aedes albopictus, by combining physicochemical analysis and behavioral assays in both an arm cage and a virtual flight arena. First, L-PDMS, when mixed with topical repellents and applied on a human skin, enhances the effect of topical repellents in reducing mosquito bites by efficiently transferring them to mosquito legs upon contact. Second, L-PDMS applied to mosquito tarsi compromises visual object tracking during flight, exerting an influence outlasting the contact. Finally, L-PDMS applied to mosquito tarsi acts as an aversive reinforcer in associative learning, making mosquitoes avoid the conditioned odor. These results uncover a multifaceted potential of L-PDMS in altering a sequence of mosquito behaviors from biting a human skin, visual object tracking following takeoff, to the response to an odor linked with L-PDMS.

Development and application of gelatin based edible coating containing cellulose nanocrystal in the storage of fresh cucumber

Cellulose nanocrystal (CNC) has attracted attention in recent times because of its unique properties. In this study, the ability of gelatin based edible coating containing CNC supplemented with glycerol was investigated to extend the shelf life of fresh cucumber (Cucumis sativum) for a storage period of 14 days. CNC was produced using chemical method and characterised using X-ray diffraction (XRD) and Fourier transform-infrared spectroscopy (FTIR). Filmogenic solution of gelatin was also produced by dissolving 10 g of gelatin powder in 100 ml of distilled water with 0, 1 and 4 g of CNC/10 g gel. The effect of two treatments alongside the control experiment was studied on some physicochemical and microbial assay were conducted. The result obtained indicate that the Derby Schrerer’s formula employed to calculate crystallite size of CNC yielded 5.9 nm. CNC- coated cucumber retained the green colour of cucumber in the first 7 days of storage. For each treatment, there were differences between the physicochemical parameters and the storage time. Weight loss, total soluble solids and titratable acidity effects were in direct correlation with the level of concentrations of the gelatin-based CNC. However, the level of retentions in ascorbic acid, flavonoids and alkaloids were higher in low treatment concentration (10000 ppm) than in the high concentration (40000 ppm) of the filmogenic coating material used. Results emanated from microbiological assays revealed that E. coli was found to be the predominant bacterial species identified in cucumber samples. No Salmonella sp. or Shigella sp. was isolated from the samples. Aspergillus sp. was found to be the most occurring fungal specie in the coated cucumber sample. Associated edible coating such as the gelatin-based CNC produced in this present research has provided reliable data for the shelf-life extension of fresh cucumber fruits at low concentration for a storage period of 2 weeks which could be used to extend the conservation period of fresh cucumber to support long period of time between harvest and transportation to distant markets.

Chemolithoautotrophic diazotrophs dominate dark nitrogen fixation in mangrove sediments.

Diazotrophic microorganisms regulate marine productivity by alleviating nitrogen limitation. So far chemolithoautotrophic bacteria are widely recognized as the principal diazotrophs in oligotrophic marine and terrestrial ecosystems. However, the contribution of chemolithoautotrophs to nitrogen fixation in organic-rich habitats remains unclear. Here, we utilized metagenomic and metatranscriptomic approaches integrated with cultivation assays to investigate the diversity, distribution, and activity of diazotrophs residing in Zhangzhou mangrove sediments. Physicochemical assays show that the studied mangrove sediments are typical carbon-rich, sulfur-rich, nitrogen-limited, and low-redox marine ecosystems. These sediments host a wide phylogenetic variety of nitrogenase genes, including groups I-III and VII-VIII. Unexpectedly diverse chemolithoautotrophic taxa including Campylobacteria, Gammaproteobacteria, Zetaproteobacteria, and Thermodesulfovibrionia are the predominant and active nitrogen fixers in the 0-18cm sediment layer. In contrast, the 18-20cm layer is dominated by active diazotrophs from the chemolithoautotrophic taxa Desulfobacterota and Halobacteriota. Further analysis of MAGs shows that the main chemolithoautotrophs can fix nitrogen by coupling the oxidation of hydrogen, reduced sulfur, and iron, with the reduction of oxygen, nitrate, and sulfur. Culture experiments further demonstrate that members of chemolithoautotrophic Campylobacteria have the nitrogen-fixing capacity driven by hydrogen and sulfur oxidation. Activity measurements confirm that the diazotrophs inhabiting mangrove sediments preferentially drain energy from diverse reduced inorganic compounds other than from organics. Overall, our results suggest that chemolithoautotrophs rather than heterotrophs are dominant nitrogen fixers in mangrove sediments. This study underscores the significance of chemolithoautotrophs in carbon-dominant ecosystems.

Customizable Three-Dimensional Printed Earring Tap for Treating Affections Caused by Aesthetic Perforations.

This work aimed to develop a three-dimensional (3D) wearable drug-loaded earring tap to treat affections caused by aesthetic perforations. The initial phase involved a combination of polymers to prepare filaments for fused deposition modeling (FDM) 3D printing using a centroid mixture design. Optimized filament compositions were used in the second phase to produce 3D printed earring taps containing the anti-inflammatory naringenin. Next, samples were assessed via physicochemical assays followed by in vitro skin permeation studies with porcine ear skin. Two filament compositions were selected for the study's second phase: one to accelerate drug release and another with slow drug dissolution. Both filaments demonstrated chemical compatibility and amorphous behavior. The use of the polymer blend to enhance printability has been confirmed by rheological analysis. The 3D devices facilitated naringenin skin penetration, improving drug recovery from the skin's most superficial layer (3D device A) or inner layers (3D device B). Furthermore, the devices significantly decreased transdermal drug delivery compared to the control containing the free drug. Thus, the resulting systems are promising for producing 3D printed earring taps with topical drug delivery and reinforcing the feasibility of patient-centered drug administration through wearable devices.

Using structural equation modeling to assess the genotoxic and mutagenic effects of heavy metal contamination in the freshwater ecosystems: A study involving Oreochromis niloticus in an urban river

Chemical pollutants represent a leading problem for aquatic ecosystems, as they can induce genetic, biochemical, and physiological changes in the species of these ecosystems, thus compromising their adaptability and survival. The Capibaribe River runs through the state of Pernambuco, located in Northeastern Brazil, and passes through areas of agricultural cultivation, densely populated cities, and industrial centers, primarily textiles. Despite its importance, few ecotoxicological studies have been conducted on its environment, and knowledge about pollution patterns and their effects on its biota is still being determined. The objective of this study was to evaluate the water quality and the damage supposed to be caused by pollutants on the DNA specimens of Nile tilapia (Oreochromis niloticus) obtained from seven strategic points of Capibaribe. Tilapia specimens and water were collected during the rainy and dry seasons from 2015 to 2017. The following characteristics were analyzed: physicochemical (six), metal concentration (seven), local pluviosity, micronuclei, and comet assay. The physicochemical and heavy metal analyses were exploratory, whereas the ecotoxicological analyses were hypothetical. To verify this hypothesis, we compared the groups of fish collected to the results of the micronuclei test and comet assay. We created a Structural Equation Model (SEM) to determine how each metal's micronuclei variables, damage index, pluviosity, and concentration were related. Our results demonstrated that the highest values for markers of genetic damage were detected at points with the highest heavy metal concentrations, especially iron, zinc, manganese, chromium, and cadmium. The SEM demonstrated that metals could explain the findings of the genotoxicity markers. Moreover, other pollutants, such as pesticides, should be considered, mainly where the river passes through rural areas. The results presented here demonstrate that the Capibaribe River has different degrees of contamination and confirm our hypothesis.

Quality Evaluation of Gentamicin Sulfate Reference Standards in Japanese Pharmacopoeia Using Hydrophilic Interaction Chromatography Combined With Tandem Mass Spectrometry.

Through the recent development of analytical technology, antibiotics quantification in the Japanese Pharmacopoeia (JP) has changed from traditional microbiological assays to physicochemical methods with high specificity and precision. However, for several multicomponent antibiotics without typical UV absorption, potency cannot be directly determined using instrumental methods such as high-performance liquid chromatography; therefore, traditional microbiological assays are still used. Gentamicin sulfate (GmS), which consists of three major components, C1, C1a, and C2, is such a typical antibiotic, and its antimicrobial potency continues to be assayed using microbiological methods in JP monographs. Introduction of a physicochemical assay for GmS is needed to help ensure its quality and quantity. This study aimed to develop quality control measures for GmS that could be complementary to quantitative assays and purity tests specified in the JP. For each gentamicin C component (C1, C2, and C1a), theoretical potencies were determined based on the quantitative relationship between purity and potency, as measured by quantitative 1H NMR and microbiological assays, respectively. Two lots of the JP reference standard (RS) were used as test samples, with the contents of each component and impurity (sisomicin and garamine) being determined using hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS). The ratios of theoretical potency for C1, C2, and C1a were 1.00, 1.21, and 1.80, respectively. The potencies of the GmS JP RSs, which were estimated based on the contents and theoretical potency of each C component, corresponded well with those determined through microbiological assays. Marked differences in impurities (%) between the two RS lots were highlighted by quantifying sisomicin and garamine. The developed analytical procedure enabled the characterization of two different JP RSs in terms of content ratio, potencies, and impurities. Novel analytical procedures useful for routine quality control of GmS were developed using HILIC-MS/MS.

Elaboration and Characterization of Dulce de Leche with Reduced Sugar Content

Dulce de leche is a South American traditional dairy product. However, due to the high sugar content, it is unsafe for consumers with special physiological conditions, such as diabetes. Light foods were developed to meet those demands whilst maintaining the sensory characteristics. The present study aimed to develop and characterize a formulation of light dulce de leche, using sweeteners to substitute sucrose, and gums as thickening and stabilizing agents. The physicochemical assays follow the AOAC manual, and the microbiological parameters were set according to MERCOSUL’s legislation. Thirty potential consumers perform the nine-point hedonic scale for product acceptance and the seven-point scale for buying intention. Compared to the traditional formulation, the product had better rheological properties, but a lighter color. All the raw materials and final products were considered safe by the recommended microbiological and toxicological standards; however, the product showed discrepancies to the physicochemical requirements. The formulation had an overall medium acceptance and low buying intention. The product had deficiencies, needing other compounds to reach the desired characteristics. Even though it leads to a more expensive final product, it is possible to adjust the product and allow access to more selective consumers or with restrictions.

High-throughput synthesis provides data for predicting molecular properties and reaction success.

The generation of attractive scaffolds for drug discovery efforts requires the expeditious synthesis of diverse analogues from readily available building blocks. This endeavor necessitates a trade-off between diversity and ease of access and is further complicated by uncertainty about the synthesizability and pharmacokinetic properties of the resulting compounds. Here, we document a platform that leverages photocatalytic N-heterocycle synthesis, high-throughput experimentation, automated purification, and physicochemical assays on 1152 discrete reactions. Together, the data generated allow rational predictions of the synthesizability of stereochemically diverse C-substituted N-saturated heterocycles with deep learning and reveal unexpected trends on the relationship between structure and properties. This study exemplifies how organic chemists can exploit state-of-the-art technologies to markedly increase throughput and confidence in the preparation of drug-like molecules.

Vitamin D3-incorporated chitosan/collagen/fibrinogen scaffolds promote angiogenesis and endothelial transition via HIF-1/IGF-1/VEGF pathways in dental pulp stem cells

Effective vascularization during wound healing remains a critical challenge in the regeneration of skin tissue. On the other hand, mesenchymal stem cell (MSC) to endothelial phenotype transition (MEnDoT) is a potential phenomenon grossly underexplored in vascularized skin tissue engineering. Vitamin D3 has a proven role in promoting MEnDoT. Hence, a D3-incorporated scaffold made with biocompatible materials such as chitosan, collagen and fibrinogen should be able to promote endothelial lineage transition in vitro for tissue engineering purposes. In this study, we developed vitamin D3 incorporated chitosan-collagen-fibrinogen (CCF-D3) scaffolds physically crosslinked under UV and conducted thorough physicochemical and biological assays on it compared to a control scaffold without vitamin D3. Our study for the first time reports the potential vascularization property of the CCF-D3 scaffold by inducing the transitions of dental pulp MSC to endothelial lineage via the HIF-1/IGF-1/VEGF pathways. MSC seeded on UV-exposed CCF-D3 scaffolds had higher cell viability and transitioned towards endothelial lineage was observed by elevated proliferative and endothelial-specific gene expressions and flow cytometric analysis of SCA-1+ antibody. The difference in VEGF-A and α-SMA expressions was also observed in the D3-CCF scaffold compared to the scaffolds without D3.