Milk-Compositional Study of Metabolites and Pathogens in the Milk of Bovine Animals Affected with Subclinical Mastitis

Bovine milk is an important food component in the human diet due to its nutrient-rich metabolites. However, bovine subclinical mastitis alters the composition and quality of milk. In present study, California mastitis testing, somatic cell count, pH, and electrical conductivity were used as confirmatory tests to detect subclinical mastitis. The primary goal was to study metabolome and identify major pathogens in cows with subclinical mastitis. In this study, 29 metabolites were detected in milk using gas chromatography–mass spectrometry. Volatile acidic compounds, such as hexanoic acid, hexadecanoic acid, lauric acid, octanoic acid, n-decanoic acid, tricosanoic acid, tetradecanoic acid, and hypogeic acid were found in milk samples, and these impart good flavor to the milk. Metaboanalyst tool was used for metabolic pathway analysis and principal component estimation. In this study, EC and pH values in milk were significantly increased (p < 0.0001), whereas fat (p < 0.04) and protein (p < 0.0002) significantly decreased in animals with subclinical mastitis in comparison to healthy animals. Staphylococcus aureus was the predominant pathogen found (n = 54), followed by Escherichia coli (n = 30). Furthermore, antibiotic sensitivity revealed that Staphylococcus aureus was more sensitive to gentamicin (79.6%), whereas Escherichia coli showed more sensitivity to doxycycline hydrochloride (80%).     

Three New Acrylic Acid Derivatives from Achillea mellifolium as Potential Inhibitors of Urease from Jack Bean and α-Glucosidase from Saccharomyces cerevisiae

(1) Background: Achillea mellifolium belongs to a highly reputed family of medicinal plants, with plant extract being used as medicine in indigenous system. However, limited data is available regarding the exploitation of the medicinal potential of isolated pure compounds from this family; (2) Methods: A whole plant extract was partitioned into fractions and on the basis of biological activity, an ethyl acetate fraction was selected for isolation of pure compounds. Isolated compounds were characterized using different spectroscopic techniques. The compounds isolated from this study were tested for their medicinal potential using in-vitro enzyme assay, coupled with in-silico studies; (3) Results: Three new acrylic acid derivatives (1–3) have been isolated from the ethyl acetate fraction of Achillea mellifolium. The characterization of these compounds (1–3) was carried out using UV/Vis, FT-IR, 1D and 2D-NMR spectroscopy (¹H-NMR, ¹³C-NMR, HMBC, NOESY) and mass spectrometry. These acrylic acid derivatives were further evaluated for their enzyme inhibition potential against urease from jack bean and α glucosidase from Saccharomyces cerevisiae, using both in-silico and in-vitro approaches. In-vitro studies showed that compound 3 has the highest inhibition against urease enzyme (IC₅₀ =10.46 ± 0.03 μΜ), followed by compound 1 and compound 2 with percent inhibition and IC₅₀ value of 16.87 ± 0.02 c and 13.71 ± 0.07 μΜ, respectively, compared to the standard (thiourea-IC₅₀ = 21.5 ± 0.01 μΜ). The investigated IC₅₀ value of compound 3 against the urease enzyme is two times lower compared to thiourea, suggesting that this compound is twice as active compared to the standard drug. On the other hand, all three compounds (1–3) revealed mild inhibition potential against α-glucosidase. In-silico molecular docking studies, in combination with MD simulations and free energy, calculations were also performed to rationalize their time evolved mode of interaction inside the active pocket. Binding energies were computed using a MMPBSA approach, and the role of individual residues to overall binding of the inhibitors inside the active pockets were also computed; (4) Conclusions: Together, these studies confirm the inhibitory potential of isolated acrylic acid derivatives against both urease and α-glucosidase enzymes; however, their inhibition potential is better for urease enzyme even when compared to the standard.     

Design and Synthesis of Some New Furan-Based Derivatives and Evaluation of In Vitro Cytotoxic Activity

New furan-based derivatives have been, designed, synthesized, and evaluated for their cytotoxic and tubulin polymerization inhibitory activities. DNA flow cytometric study of pyridine carbohydrazide 4 and N-phenyl triazinone 7 demonstrated G₂/M phase cell cycle disruptions. Accumulation of cells in the pre-G1 phase and positive annexin V/PI staining, which may be caused by degeneration or fragmentation of the genetic components, suggested that cell death occurs via an apoptotic cascade. Furthermore, compounds 4 and 7 had a strong pro-apoptotic impact through inducing the intrinsic mitochondrial mechanism of apoptosis. This mechanistic route was verified by an ELISA experiment that indicated a considerable rise in the levels of p53 and Bax and a drop in the level of Bcl-2 when compared with the control.     

Design,Synthesis and Cytotoxic Activity Evaluation of Newly Synthesized Amides-Based TMP Moiety as Potential Anticancer Agents over HepG2 Cells

A novel series of amides based TMP moiety was designed, synthesized and evaluated for their antiproliferative as well as enzyme inhibition activity. Compounds 6a and 6b showed remarkable cytotoxic activity against HepG2 cells with IC₅₀ values 0.65 and 0.92 μM, respectively compared with SAHA and CA-4 as reference compounds. In addition, compound 6a demonstrated good HDAC-tubulin dual inhibition activity as it showed better HDAC activity as well as anti-tubulin activity. Moreover, compound 6a exhibited G2/M phase arrest and pre-G1 apoptosis as demonstrated by cell cycle analysis and Annexin V assays. Further apoptosis studies demonstrated that compound 6a boosted the level of caspase 3/7. Caspase 3/7 activation and apoptosis induction were evidenced by decrease in mitochondrial permeability suggesting that activation of caspase 3/7 may occur via mitochondrial apoptotic pathway.     

Covalent immobilization of Candida antarctica lipase B on nanopolystyrene and its application to microwave-assisted esterification

Nanopolystyrene was used as a solid support for the covalent immobilization of Candida antarctica lipase B (CalB) using the photoreactive reagent 1-fluoro-2-nitro-4-azido benzene (FNAB) as a coupling reagent. The obtained derivative was then used as a biocatalyst in a microwave assisted esterification experiment. Factors such as contact time, pH, and enzyme concentration were investigated during immobilization. The hydrolytic activity, thermal, and operational stability of immobilized-CalB were determined. The maximum immobilized yield (218 μg/mg support) obtained at pH 6.8 exhibited optimum hydrolytic activity (4.42 × 10³ mU p-nitrophenol/min). The thermal stability of CalB improved significantly when it was immobilized at pH 10, however, the immobilized yield was very low (93.6 μg/mg support). The immobilized-CalB prepared at pH 6.8 and pH 10 retained 50% of its initial activity after incubation periods of 14 and 16 h, respectively, at 60 ℃. The operational stability was investigated for the microwave assisted esterification of oleic acid with methanol. Immobilized-CalB retained 50% of its initial activity after 15 batch cycles in the microwave-assisted esterification. The esterification time was notably reduced under microwave irradiation. The combined use of a biocatalyst and microwave heating is thus an alternative total green synthesis process.     

1,3-Diyl trapping reactions. Fundamental investigations with application to the synthesis of linearly fused tricyclopentanoids

A comparison of the inter- and intramolecular diyl trapping routes to linearly fused tricyclopentanoids is presented. In addition, several of the factors which are responsible for the stereoselectivity which is associated with the intramolecular process are examined and it is concluded that conformational rather than electronic (secondary orbital) factors play the dominant role. It is shown that gem methyl groups located on the acyclic chain which joins the diyl and diylophile (in reference to 32 and 35, but not to 47) have no practical effect upon the outcome of the trapping reaction. The intramolecular process is stereospecific with respect to diylophile geometry, and highly stereoselective with respect to the ring junction stereochemistry. Finally, an abortive attempt to synthesize the marine natural product Δ⁹⁽¹²)-capnellene (19) as well as a successful synthesis of the mold metabolite d,l- hirsutene (18) is presented.     

Spectrofluorimetric determination of fexofenadine hydrochloride in pharmaceutical preparation using silver nanoparticles

A novel sensitive fluorimetric method was investigated for the assay of fexofenadine hydrochloride (FEX) using silver nanoparticles (NPs) as a fluorescence probe. The NPs, which were prepared by chemical reduction of silver nitrate with sodium borohydride (reducing agent) in aqueous solution (without organic stabilizers) were water soluble, stable and had narrow emission band. The addition of drug to NPs solution caused considerable quenching of the emission band of silver NPs, which was likely due to the complexation of the drug to silver NPs. Under the optimum conditions, the quenched fluorescence (FL) intensity was linear with the concentration of FEX in the range of 1 × 10^?7 to 2.5 × 10^?5 mol L^?1 (0.9985) with a detection limit of 1.2 × 10^?8 mol L^?1. The quenching mechanism of the studied drug on the emission band of silver NPs was explained by Stern–Volmer law. The developed method was applied to FEX determination in a pharmaceutical formulation (allegra tablets) and biological fluids (human serum and urine).     

Spectro-spatial analyses of a nonlinear metamaterial with multiple nonlinear local resonators

Nonlinear Dynamics - Recent focus has been given to spectro-spatial analysis of nonlinear metamaterials since they can predict interesting nonlinear phenomena not accessible by spectral analysis...     

南极假丝酵母在纳米聚苯乙烯上的共价固载及其在微波辅助酯化反应中的应用（英文）

Nanopolystyrene was used as a solid support for the covalent immobilization of Candida antarctica lipase B(Cal B) using the photoreactive reagent 1‐fluoro‐2‐nitro‐4‐azido benzene(FNAB) as a cou‐pling reagent. The obtained derivative was then used as a biocatalyst in a microwave assisted ester‐ification experiment. Factors such as contact time, pH, and enzyme concentration were investigated during immobilization. The hydrolytic activity, thermal, and operational stability of immobi‐lized‐Cal B were determined. The maximum immobilized yield(218 μg/mg support) obtained at pH 6.8 exhibited optimum hydrolytic activity(4.42 × 103 mU p‐nitrophenol/min). The thermal stability of Cal B improved significantly when it was immobilized at pH 10, however, the immobilized yield was very low(93.6 μg/mg support). The immobilized‐Cal B prepared at pH 6.8 and pH 10 retained 50% of its initial activity after incubation periods of 14 and 16 h, respectively, at 60 °C. The opera‐tional stability was investigated for the microwave assisted esterification of oleic acid with metha‐nol. Immobilized‐Cal B retained 50% of its initial activity after 15 batch cycles in the micro‐wave‐assisted esterification. The esterification time was notably reduced under microwave irradia‐tion. The combined use of a biocatalyst and microwave heating is thus an alternative total green synthesis process.