Preparation Of Extracts Research Articles

Effect of The Water Origin on The Biological Properties of Sage (Salvia officinalis L.) Aqueous Extracts

Plant extracts, due to the high content of various biologically active compounds, have found their wide application in the foods, pharmaceutical industries, and agricultures, for the production of valuable and useful bioproducts. The aqueous plant extracts are most often used. The effect of the water origin used in their preparation on the extract’s composition has not been studied enough, which limits their use. The study was aimed to compare the biological effect of sage (Salvia officinalis L.) extracts prepared using water of different origins (distilled, tap, spring, bottled). The preparation of aqueous extracts was carried out in two ways, differing in the extraction temperature. In the first method, boiling water was used to prepare the extract; in the second, cooled boiled water was used. The sage leaves mass for extract preparation was 1.65 and 3.30 g. The biological activity of the extracts was assessed by testing them for radish germination. It was shown the sage leaves extracts prepared using various waters origin can differ by one and half times in their allelopathic effect on radish seeds. It was proved the qualitative composition and mass of the substance extracted from the plant depend on the water origin used for extracts preparation. Thus, using the same amount of plant material to prepare extracts, you can increase their biological activity by fifty percent with the right choice of water.

Formulation and evaluation of herbal gel containing ethanolic extract of Momordica charantia against socransky’s periodontal pathogens of the oral cavity-an in vitro study

Background: Periodontitis is a multifaceted disease initiated by periodontal pathogens which when left untreated results in inflammation of the supporting tissues of the teeth, resorption of alveolar bone, loss of the periodontal ligament attachment, periodontal pocket formation and/or recession of the gingiva. Hence, the current study aimed at evaluating the efficacy of Momordica charantia leaves extract on Socransky’s periodontal pathogens and formulating a herbal gel for subgingival local drug delivery for the potential treatment of moderate periodontitis. Methods: M. charantia leaves were authenticated from a recognized taxonomist. They were coarsely powdered following which an ethanol-based extract preparation was done. The extract obtained was then assessed for minimum inhibitory concentration, minimum bactericidal concentration followed by gel formulation which was then subjected to time kill assay and zone of inhibition on periodontal pathogens, the efficacy of which was comparatively evaluated against 0.12% chlorhexidine (CHX) gluconate. Kruskal-Wallis Test was employed with the statistical significance set at p≤0.05. Results: The concentration of M. charantia ethanolic extract against periodontal pathogens was determined to be 1 gm/100 ml and a statistically significant difference was found in the effectiveness between the gel so formulated and CHX. Conclusion: The antibacterial activity was evident in the ethanolic extract of M. charantia which when used to formulate a herbal gel against anaerobic periodontal pathogens demonstrated efficacy comparable to that of CHX.

Differences of in vitro immune responses between patent and pre-patent Litomosoides sigmodontis-infected mice are independent of the filarial antigenic stimulus used.

Lymphatic filariasis and onchocerciasis are neglected tropical diseases and cause significant public health problems in endemic countries, especially in sub-Saharan Africa. Since the human parasites are not viable in immune-competent mice, animal models have been developed, among them Litomosoides sigmodontis which permits a complete life cycle in BALB/c mice, including the development of patent infections with circulating microfilariae (Mf, the worm's offspring). To investigate the immunomodulatory properties of helminths in vitro, antigenic extracts can be prepared from different life cycle stages of the L. sigmodontis model, including adult worms, but the methods to prepare these antigens differ between research groups. This study analyzed whether different centrifugation methods during the preparation of an antigenic extract, the gender of used worms, or the different fractions (soluble or pellet) altered filarial-specific CD4+ Tcell responses. These cells were isolated from pre-patent or patent/chronic infected mice, hence those without and those with Mf, respectively. Ex vivo immune responses were compared at these two different time points of the infection as well as the parasitic parameters. Worm burden and cell infiltration were elevated in the thoracic cavity (TC) and draining mediastinal lymph nodes at the pre-patent stage. Within the TC, eosinophils were significantly up-regulated at the earlier time point of infection which was further reflected by the eosinophil-related eotaxin-1 levels. Regarding the production of cytokines by re-stimulated CD4+ T cells in the presence of different antigen preparations, cytokine levels were comparable for all used extracts. Our data show that immune responses differ between pre-patent and patent filarial infection, but not in response to the different antigenic extracts themselves.

NANOPORE-BASED INTRA-OPERATIVE CLASSIfiCATION OF CENTRAL NERVOUS SYSTEM TUMOURS IN THE NHS

Abstract AIMS The 2021 WHO classification of CNS tumours necessitates the use of molecular testing in order to reach a confident diagnosis of many tumour types. At present, this is achieved using array-based technology delivered by a centralised model. This approach has inherent delays that mean it is often several weeks before the treating clinical team is informed of diagnosis, delaying the start of adjuvant therapy and causing significant distress for patients. As such, there is an urgent need to rethink this pathway and improve time to diagnosis. Nanopore longread sequencing technology has the potential to enable a paradigm shift in diagnostic and therapeutic pathways, offering rapid, comprehensive molecular diagnosis that can be performed in the intra-operative setting. We aimed to test the hypothesis that intra-operative methylation-based classification can be performed using nanopore sequencing and thereby expediently inform surgical and oncological decision-making. METHOD Oxford Nanopore Technology (ONT) is a native-strand, long-read sequencing platform that can perform copy- number profiling along with parallel single-gene methylation, structural variant, mutational and methylation analyses. Capital and consumable costs are relatively low, and testing can be performed in the local setting on single samples. We have optimised DNA extraction and library preparation for rapid tissue processing. We have developed a robust bioinformatic pipeline which is able to produce intraoperative classification within minutes of sequencing and full molecular profiling after 24 hours. RESULTS We demonstrate the feasibility of using ONT to deliver intra-operative methylation-based classification of CNS tumours in ‘realtime’ within an NHS hospital setting and will discuss the challenges of implementing this testing and the opportunities to inform neuro-oncological practice. CONCLUSION By giving a summary of our novel protocol of rapid DNA extraction, library preparation and bioinformatic analysis, we will show that this approach can revolutionise the diagnosis of brain tumours and has the potential for rapid adoption by non-expert laboratories.

Quantifying allergenic proteins using antibody-based methods or liquid chromatography-mass spectrometry/mass spectrometry: A review about the influence of food matrix, extraction, and sample preparation.

Accurate quantification of allergens in food is crucial for ensuring consumer safety. Pretreatment steps directly affect accuracy and efficiency of allergen quantification. We systematically reviewed the latest advances in pretreatment steps for antibody-based methods and liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) protein quantification methods in food. For antibody-based methods, the effects induced by food matrix like decreased allergen solubility, epitope masking, and nonspecific binding are of the upmost importance. To mitigate interference from the matrix, effective and proper extraction can be used to obtain the target allergens with a high protein concentration and necessary epitope exposure. Removal of interfering substances, extraction systems (buffers and additives), assistive technologies, and commercial kits were discussed. About LC-MS/MS quantification, the preparation of the target peptides is the crucial step that significantly affects the efficiency and results obtained from the MS detector. The advantages and limitations of each method for pre-purification, enzymatic digestion, and peptide desalting were compared. Additionally, the application characteristics of microfluidic-based pretreatment devices were illustrated to improve the convenience and efficiency of quantification. A promising research direction is the targeted development of pretreatment methods for complex food matrices, such as lipid-based and carbohydrate-based matrices.

Nanopore ultra-long sequencing and adaptive sampling spur plant complete telomere-to-telomere genome assembly

The pursuit of complete telomere-to-telomere (T2T) genome assembly in plants, challenged by genomic complexity, has been advanced by Oxford Nanopore Technologies (ONT), which offers ultra-long, real-time sequencing. Despite its promise, sequencing length and gap filling remain significant challenges. This study optimized DNA extraction and library preparation, achieving DNA lengths exceeding 485 kb; average N50 read lengths of 80.57 kb, reaching up to 440 kb; and maximum reads of 5.83 Mb. Importantly, we demonstrated that combining ultra-long sequencing and adaptive sampling can effectively fill gaps during assembly, evidenced by successfully filling the remaining gaps of a near-complete Arabidopsis genome assembly and resolving the sequence of an unknown telomeric region in watermelon genome. Collectively, our strategies improve the feasibility of complete T2T genomic assemblies across various plant species, enhancing genome-based research in diverse fields.

Ion chromatography coupled with optical emission spectrometry (IC-ICP-OES) methodology for the analysis of inositol phosphates in food and feed

This study presents the development of ion chromatography coupled with inductively coupled plasma optical emission spectrometry (IC-ICP-OES) for the simultaneous determination and quantification of inositol phosphates (InsPx). Using a CarboPac PA100 column with a nitric acid-water gradient, 28 InsPx isomers (InsP6 to InsP2) were separated within 33 min. The method eliminates baseline drift and post-column derivatization thereby simplifying detection and quantification. It achieves low detection limits of 63 μg/L P across a range of 63–3200 μg/L P. Various extraction and sample preparation methods for food and feed matrices were tested, including acidic and alkaline agents, C18 SPE and spin concentrators. The analysis shows intra-day and intra-laboratory reproducibility with deviations smaller than 1 % for standard solutions and under 4 % for feed samples (80 % recovery rate of phytate). This methodology is applicable to explore enzymatic degradation pathways and the analysis of InsPx in complex food and animal feed matrices.

Exploring Antimicrobial Phytocompounds in Tanacetum dolicophyllum: A GC-MS Profiling Approach.

Tanacetum dolicophyllum, known locally as tansies, exhibitng a plethora of challenging and noteworthy pharmacological properties. Aerial parts of T. dolicophyllum were utilized for extract preparation, and then their in-vitro antimicrobial activity was assessed against various organisms employing both the well diffusion method and MIC assay. The ethyl acetate extract of T. dolicophyllum was analyzed using gas chromatography-mass spectrometry (GC-MS) investigation to determine phytocomponents and investigate their antimicrobial activity. The result demonstrated that ethyl acetate extract exhibited good antimicrobial activity. The extract showed the highest efficacy against Klebsiella pneumoniae, Streptococcus agalactiae, S. pyogenes, Escherichia coli, and Candida albicans. Analysis revealed the presence of 37 components in the T. dolicophyllum extract, as determined by GC-MS investigation. The most abundant compounds include pentacosane, constituting 17.31 of the peak area% with a retention time (RT) of 24.603, and molecular formula C25H52; 2(3H)-Furanone, dihydro-3-[(7-methoxy1,3-benzodioxol-5-yl)methyl]-4-[(3,4,5-trimethoxyphenyl)methyl]- accounting for 6.43 peak area% with a RT of 31.829 and, molecular formula C23H26O8; 4-(1,3-benzodioxol-5-ylmethyl)dihydro-3-[(3,4,5-trimethoxyphenyl)methyl]- representing 5.76 peak area%, RT 28.409 and, molecular formula C22H24O7; Yangambin present at 5.44% of peak area, with an RT of 29.513 and, molecular formula C24H30O8; Chrysantenyl 2-methuylbutanoate constituting 4.60 of peak area%, RT 18.825, and molecular formula C15H24O2; Phytyl tetradecanoate with 3.22 area%, RT 31.284, and formula C34H66O2; and Tricosane accounting for 1.44 peak area%, with a RT of 18.228, and molecular formula C23H48. These compounds exhibit various medical potentials, suggesting the ethyl acetate extract of T. dolicophyllum may have diverse pharmacological applications.

Extract Preparation of Waste Lady Finger Caps Using Ethanol, Generation of Extract’s Layers on Copper Through Drop Casting Without and with NiO Nanoparticles, and Study of their Corrosion Performances in Saline Water

Extract Preparation of Waste Lady Finger Caps Using Ethanol, Generation of Extract’s Layers on Copper Through Drop Casting Without and with NiO Nanoparticles, and Study of their Corrosion Performances in Saline Water

GC–MS metabolomics of French lettuce (Lactuca Sativa L. var capitata) leaves exposed to bisphenol A via the hydroponic media

IntroductionBisphenol A (BPA), an organic compound used to produce polycarbonate plastics and epoxy resins, has become a ubiquitous contaminant due to its high-volume production and constant release to the environment. Plant metabolomics can trace the stress effects induced by environmental contaminants to the variation of specific metabolites, making it an alternative way to study pollutants toxicity to plants. Nevertheless, there is an important knowledge gap in metabolomics applications in this area.ObjectiveEvaluate the influence of BPA in French lettuce (Lactuca Sativa L. var capitata) leaves metabolic profile by gas chromatography coupled to mass spectrometry (GC–MS) using a hydroponic system.MethodsLettuces were cultivated in the laboratory to minimize biological variation and were analyzed 55 days after sowing (considered the plant’s adult stage). Hexanoic and methanolic extracts with and without derivatization were prepared for each sample and analyzed by GC–MS.ResultsThe highest number of metabolites was obtained from the hexanoic extract, followed by the derivatized methanolic extract. Although no physical differences were observed between control and contaminated lettuce leaves, the multivariate analysis determined a statistically significant difference between their metabolic profiles. Pathway analysis of the most affected metabolites showed that galactose metabolism, starch and fructose metabolism and steroid biosynthesis were significantly affected by BPA exposure.ConclusionsThe preparation of different extracts from the same sample permitted the determination of metabolites with different physicochemical properties. BPA alters the leaves energy and membrane metabolism, plant growth could be affected at higher concentrations and exposition times.

Identification of 69 magnesium-free chlorophyll derivatives in decarboxylated extracts of medical Cannabis chemovars using ESI-LC-HRMS/MS

The Cannabis plant contains many groups of bioactive metabolites with potential pharmacological properties. These properties have mainly been attributed to cannabinoids and terpenoids, with other groups receiving little attention. The usefulness of the abundant group of chlorophyll derivatives (CDs) was illustrated in various applications, but little is known regarding their presence and significance in Cannabis. We hypothesized that the heating accompanying the process extract preparation would result in a pool of CDs that have lost the central magnesium (Mg) ion. Herein, we introduce a liquid chromatography-tandem mass spectrometry approach for separating, identifying, and quantifying Mg-free CDs in Cannabis extracts after decarboxylation. We assessed 14 Cannabis cultivars representing the four types of chemovars and identified 69 distinct Mg-free CDs. These were separated into five families: Pheophorbide a, Pheophorbide b, Pheophytin a, Pheophytin b, and Purpurin 18. The structure of the Mg-free CDs was determined through their MS/MS fragmentation spectra as phytylated derivatives compared to dephytylated ones, and then further to pyroderivatives, alkylated and oxidized CDs. Substantial variation was found between the four different chemovar types and within different cultivars of the same chemovar, with the family of Pheophytin a, the most prevalent in all extracts. Type III chemovar high-cannabidiol plants contained significantly higher amounts of Mg-free CDs. These differences in abundance may result in variations in the therapeutic effects of plants that are considered similar according to their cannabinoid profiles.

Ethnobotanical diversity, phytochemical screening and biological evaluation of selected medicinal mushrooms species

The current study was aimed to investigate the ethno-pharmacology, diversity, biological potentials of different mushrooms species from District Swabi, Pakistan. Ethno-pharmacological data was collected from 73 respondents based on interview, group discussion and field visits. Quantitative analysis such as informant consent factor (ICF), Use value (UV), Relative Frequency of Citation (RFC) were done. Morphological identification of mushrooms, extract preparation, phytochemical screening and statistical analysis of data was conducted. Phytochemical analysis of six mushroom species revealed that saponins, alkaloids, glycosides, flavonoids and phenols were present in six selected mushrooms, while tannin and steroids (−) present in Agaricus arvensis. In Morchella conica only steroids (−) was absent, Qunion (−) in Polyporus tuberaster, Pleurotus ostreatus and Omphalotus olearius were found absent. Moisture contents (10.17 ± 1.27) and crude fats (5.26 ± 1.50) present in Omphalotus olearius. Highest content of crude fibers (26.57 ± 0.86) and carbohydrates (62.18 ± 0.73) were found in Pleurotus ostreatus. Highest ash contents (12.62 ± 0.46) were reported in Polyporus tuberaster. The respondents’ consensus factor values ranged from 1.66 to 61.74 in which highest ICF value was 61.74 while the lowest was 1.66. The high RFC was recorded forAgaricus (0.67) while lowest RFC was observed in Clitocybe frondasa (0.05). The results of analgesic activity were compared to the diclofenac (Standard drug). The extracts of aqueous and methanolic extract were most active at the level of (400 mg/kg). As antipyretic agents, methanolic and aqueous extracts of Agaricus species were analyzed for anti-inflammatory activity compare to diclofenac (Standard drug) where the aqueous (200 mg/kg) and methanolic (400 mg/kg) were found to be very active. Finally, mushrooms with high (ICF and UV) were further investigated for phytochemicals and pharmacological analysis which revealed significant biological potentials. This provides basic ground information which can be used to explore novel mushroom-based medicines for the treatment of various diseases. The commercial cultivation of mushrooms represents a promising opportunity to leverage their diverse health benefits and nutritional properties. By prioritizing research and cultivation methods while educating the public, the potential of mushrooms as vital components of both nutrition and medicine can be significantly realized, impacting health outcomes and contributing to sustainable agricultural practices.

Type I Kounis syndrome in a young woman without chest pain: a case report

BackgroundKounis syndrome is defined as the concurrence of acute coronary syndromes in the setting of allergic or anaphylactic reactions. It primarily affects men aged 40–70 years and is often associated with chest pain. This syndrome is often unrecognized and undiagnosed in clinical practice due to a low level of awareness. Herein, we present a case of type I Kounis syndrome in a young woman without chest pain.Case presentationA 28-year-old Japanese woman with a history of atopic dermatitis received a glycyrrhizin, glutathione, and neurotropin preparation (a preparation of inflamed skin extract from rabbits inoculated with vaccinia virus) at a dermatology clinic to treat pruritus caused by atopic dermatitis. Immediately after the administration, the patient developed abdominal pain and generalized body wheals. The patient was diagnosed with anaphylaxis and was transported to our hospital. She had no chest pain on arrival at our hospital; however, a 12-lead electrocardiogram showed ST elevation in leads I, aVL, V2, and V3, and an echocardiogram showed decreased wall motion in the anterior and lateral walls of the left ventricle. Sublingual nitroglycerin administration improved ST-segment elevation and left ventricular wall motion abnormalities. The patient underwent emergency coronary angiography, which revealed no significant stenosis, and was diagnosed with type I Kounis syndrome.ConclusionKounis syndrome without chest pain is rare in young women. Since it can be fatal in cases with severe allergic symptoms such as anaphylaxis, the possibility of concurrent acute coronary syndrome should be considered when treating systemic allergic reactions, regardless of age, sex, or the presence or absence of chest symptoms.

HPLC-DAD method validation for quantification of dehydroabietic acid and abietic acid in oral spray containing Pinus merkusii heartwood extract and its antibacterial effects on clinically isolated Streptococcus mutans

AbstractPinus merkusii Jungh & De Vries. has become increasingly gathered more attention from researchers because the plant has a range of folk medicinal uses. Heartwood plant is the major source of dehydroabietic acid (DHAA) and abietic acid (AA), which possesses several medicinal properties, such as antiviral, antimicrobial, antiobesity and anti-inflammatory. The research proposed herein a low-cost, fast, specific, uncomplicated, sensitive, precise reverse-phase high-performance liquid chromatography (RP-HPLC). This method was conducted and validated for evaluating an amount of DHAA and AA in ethanol extract and oral spray containing P. merkusii heartwood extract. Additionally, stability and antimicrobial activities against clinically isolated Streptococcus mutans of the oral spray were determined. The separation was achieved on Pursuit 200Å PFP column, 150 × 4.6 mm, particles of 3 µm with a flow rate of 1.0 mL min−1. Methanol and water (70:30 v/v) were used as eluent with an isocratic mode and sample analysis volume was set at 10 µL, at a detection wavelength of 210 and 245 nm. The developed HPLC method for analysis of DHAA and AA showed good linearity with correlation coefficients equal to 1. Moreover, other validation parameters, comprised of accuracy, precision, specificity and detection and quantitation limits of this method displayed excellent reliability, validity and sensitivity. This method could be an interesting alternative for quantitative measurement of P. merkusii heartwood extract, oral spray formulation and other P. merkusii heartwood extract preparations. The result from antibacterial tests suggested that the oral spray containing P. merkusii heartwood extract is able to inhibit the oral pathogens causing dental caries. The oral spray decreased S. mutans population size by about 0.5–2 Log CFU mL−1 at 1–4 h and complete elimination of all bacteria strains within 24 h. This study provides validity for using P. merkusii heartwood extract as an alternative for preventing and treating oral infectious diseases.

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.

Novel Synthesis and Biophysical Characterization of Zinc Oxide Nanoparticles Using Virgin Coconut Oil.

One of the primary concerns in the field of green synthesis of nanoparticles (NPs) utilizing plant materials is the scarcity of high purity, challenges in achieving large-scale production, and limited global accessibility. Hygienic preparation and safe storage of plant extracts are also considerable challenges in this field. So, an investigation was started to overcome these limitations. Virgin coconut oil (VCO) in its purest form is available commercially all over the world. Also, it has high medicinal value with excellent biomedical applications. Very limited work has been reported on oils as bio reducers and stabilizers. In those reports, they used a few chemicals as mediators in the processes of synthesis and cleaning. So, to the best of our knowledge, for the first time, zinc oxide (ZnO) NPs were synthesized using VCO as a reducing and capping agent with zero chemical mediators. A comprehensive investigation of the structural, microstructural, and optical properties was reported. X-ray diffraction confirms the formation of VCO-ZnO NPs with an average crystallite size of 32.81 nm in a hexagonal structure. UV characteristics confirm quantum confinement through a well-defined SPR near 223 nm with fwhm of 67 nm and a direct band gap at 3.96 eV. FTIR reveals the capping of VCO through carboxylic functional groups, particularly the -COO- group of coconut oil at 1770 cm-1 with a shift of about 30 cm-1 compared to plain VCO. TEM confirms the polycrystalline nature with nearly spherical and 10-22 nm particle size. The zeta potential of -15.4 ± 5.0 mV signifies the stability and antiagglomeration properties. FESEM with EDS results confirms morphological excellence, the purity level of synthesized NPs (99.5%), and the prominent scalability of NPs (84.38% yield). Finally, as-synthesized VCO-ZnO NPs showed very good antioxidant (IC50 78.991, 51.464, and 4.677 μg/mL in DPPH, ABTS, and FRAP assays, respectively), anti-inflammatory (IC50 22.42 μg/mL in protein denaturation), antimicrobial (MIC 0.156 mg/mL for Pseudomonas and 0.316 mg/mL for S. aureus), and antidiabetic properties (IC50 88.45 and 147.67 μg/mL for α-amylase and α-glucosidase assays, respectively).

Enhancing the biological activities and altering the chemical structure of clove aqueous extract prepared with plasma-activated water as a novel technique

This investigation assesses the effect of plasma-activated water (PAW) as a solvent during the preparation of clove aqueous extract. PAW was generated by surface dielectric barrier discharge (SDBD) plasma device for 5, 10, and 15 min, respectively. The conventional method of maceration was used as a control group. We evaluated the biological activity and physicochemical characteristics of clove extracts. In the color assay, the clove extract produced by PAW showed a slightly higher color parameter than the control extract. The Fourier transform infrared (FTIR) results demonstrated several higher absorption intensities at specific regions in the PAW-clove extract structure. Liquid chromatography-mass spectrometry (LC-MS) analysis of the PAW extracts showed more bioactive components than the maceration extract. The clove extract prepared by the PAW -5 min group had the highest total phenolic content (TPC) at 821.6 mg GAE/g, the highest free radical scavenging activity (DPPH) and ferric-reducing antioxidant power (FRAP) compared to other groups. These alterations could be due to reactive oxygen-nitrogen species (RONS) generated in PAW, which can modify the physicochemical properties of clove. The findings indicated that PAW is a unique extraction solvent that improves the extraction of phenolic compounds from clove.

Exploring the potential of chitosan from royal shrimp waste for elaboration of chitosan/bioglass biocomposite: Characterization and “in vitro” bioactivity

Exploiting royal shrimp waste to produce value-added biocomposites offers environmental and therapeutic benefits. This study proposes biocomposites based on chitosan and bioglass, using shrimp waste as the chitosan source. Chitin extraction and chitosan preparation were characterized using various analytical techniques. The waste composition revealed 24 % chitin, convertible to chitosan, with shells containing 77.33-ppm calcium. (X-ray diffraction) XRD analysis showed crystallinity index of 54.71 % for chitin and 49.14 % for chitosan. Thermal analysis indicated degradation rates of 326 °C and 322 °C, respectively. The degree of deacetylation of chitosan was 97.08 % determined by proton nuclear magnetic resonance (1H-NMR) analysis, with an intrinsic viscosity of 498 mL.g−1 and molar mass of 101,720 g/mol, showing improved solubility in 0.3 % acetic acid. Royal chitosan (CHR) was combined with bioglass (BG) via freeze-drying to create a CHR/BG biocomposite for bone surgery applications. The bioactivity of the CHR/BG was tested in simulated body fluid (SBF), revealing a biologically active apatite layer on its surface. Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES) analysis confirmed enhanced bioactivity of the CHR/BG compared to commercial chitosan. The CHR/BG biocomposite demonstrated excellent apatite formation, validated by Scanning Electron Microscopy (SEM), highlighting its potential in bone surgery.

Ilex paraguariensis Extracts: A Source of Bioelements and Biologically Active Compounds for Food Supplements

Ilex paraguariensis, commonly known as yerba mate, is a plant belonging to the holly genus Ilex and the Aquifoliaceae family, indigenous to South America, and is used for the production of yerba mate. Yerba mate is renowned for its abundance of essential nutrients and bioactive compounds. Based on test results, it can be assumed that the selection of raw material for the preparation of extracts as well as the extraction method significantly influence the final content of biologically active compounds in the extracts. Consequently, this variability impacts the ultimate concentration of biologically active substances within the end product, potentially influencing human consumption. The present study aimed to quantify and compare the content of selected biological active compounds in supplements and products containing I. paraguariensis extracts, along with organic yerba mate dried through a smoke-free process, available in the European market (P-1–P-10). The evaluation focused on antioxidant substances such as neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, 4-feruloylquinic acid, isochlorogenic acid, rutoside astragalin, and caffeine. Additionally, the concentration of specific macro and trace elements was ascertained. The antioxidant compound makeup differs between methanol-extracted samples and aqueous extracts. In both cases, methanol extracts, particularly those in instant and traditional herb forms, showed the highest content of organic compounds with antioxidant properties (such as phenolic compounds and caffeine). The highest content of chlorogenic acid was detected in both methanol (14.7412 mg/g d.w.) and water (8.3120 mg/g d.w.) extracts in product P-4. The caffeic acid content ranged from 0.1491 mg/g d.w. to 1.7938 mg/g d.w. in methanol extracts and from 0.0760 mg/g d.w. to 0.4892 mg/g d.w. in water extracts. The neochlorogenic acid content ranged from 2.6869 to 23.9750 mg/g d.w. in ethanol extracts and from 0.4529 to 10.2299 mg/g d.w. in water extracts. Therefore, the traditional preparation of yerba mate as a water infusion does not fully exploit the raw material’s potential. Among the tested products, only the dietary supplement in capsule form contained protocatechuic acid, which was not present in any other tested products. Conversely, compounds characteristic of yerba mate found in other preparations were absent in this supplement. The caffeine content was also the lowest in this product. The determined content of active substances did not consistently match the declarations made by producers if stated on the packaging.

Establishing a High-Yield Chloroplast Cell-Free System for Prototyping Genetic Parts.

Plastid engineering offers the potential to carry multigene traits in plants; however, it requires reliable genetic parts to balance expression. The difficulty of chloroplast transformation and slow plant growth makes it challenging to build plants just to characterize genetic parts. To address these limitations, we developed a high-yield cell-free system from Nicotiana tabacum chloroplast extracts for prototyping genetic parts. Our cell-free system uses combined transcription and translation driven by T7 RNA polymerase and works with plasmid or linear template DNA. To develop our system, we optimized lysis, extract preparation procedures (e.g., runoff reaction, centrifugation, and dialysis), and the physiochemical reaction conditions. Our cell-free system can synthesize 34 ± 1 μg/mL luciferase in batch reactions and 60 ± 4 μg/mL in semicontinuous reactions. We apply our batch reaction system to test a library of 103 ribosome binding site (RBS) variants and rank them based on cell-free gene expression. We observe a 1300-fold dynamic range of luciferase expression when normalized by maximum mRNA expression, as assessed by the malachite green aptamer. We also find that the observed normalized gene expression in chloroplast extracts and the predictions made by the RBS Calculator are correlated. We anticipate that chloroplast cell-free systems will increase the speed and reliability of building genetic systems in plant chloroplasts for diverse applications.