[12] Determination of total protein

Growing interest has been shown in hypoglycaemic medications made from natural substances, notably those produced from plants. Flavonoids are phenolic compounds that exist in nature and have a broad range of biological activities and beneficial effects in the therapy of metabolic disorders.

The research aimed to purify apigenin (AP) from parsley leaves and study its effect on the levels of some important serum biomarkers (metabolic, oxidant and antioxidant) in rats induced diabetes in comparison with crude parsley leaves extract (PLE).

The study's findings demonstrated that apigenin is successfully purified from parsley leaves with a high yield and purity. Based on the outcomes of the experiment, the levels of all blood biomarkers showed improvement in diabetic treated group with either PLE or AP. Notably, PLE was more effective than AP in the enhancement of metabolic parameters. Considering the oxidant biomarkers (xanthine oxidase, peroxynitrite anion radical and malondialdehyde), AP had a stronger ability to reduce these harmful oxidants than PLE. Furthermore, the data of antioxidant activity showed that both compounds were able to improve the antioxidant enzyme activity. Noteworthy, AP increases the activity of glutathion-S-transferase (GST) and aminoacylase-1 (ACY1) more than PLE. Besides, PLE displays more effective in the activity of superoxide dismutase (SOD) and paraxonase (PON1). In the manner of non-antioxidant activity AP enhanced the level of metallothionein (MT), overall antioxidant activity (AOA), copper (Cu) more than PLE while, PLE showed stronger ability to improve zinc (Zn) levels.

Echinodorus macrophyllus (Kunth.) Micheli (Alismataceae), known as chapéu-de-couro in Brazil, is popularly used to treat inflammatory diseases. We have previously demonstrated a significant reduction in the acute inflammation for the aqueous extract of E. macrophyllus (AEEm) and its ethanolic fraction (Fr20) and described that hydroxycinnamoyl derivatives present in SF1 (Fr20 subfraction) showed higher anti-inflammatory properties by mechanisms that include a reduction of TNF-α, IL-1β, CKCL1/KC, LTB4, and PGE2 levels in exudate.

This work describes the acute toxicological effect of SF1 subfraction on SW mice treated orally for five days in the air pouch model by evaluating the hematological and biochemical determinations on the blood samples; the relative organ weight and its histopathological analysis; the liver genotoxicity assessment and the activity of liver enzymes from xenobiotic metabolism.

Fr20 was earlier fractionated on the Sephadex LH-20 column, yielding mainly four subfractions, including SF1. The SF1 toxicity was evaluated in mice challenged with carrageenan on the air pouch inflammation model and orally treated for five days. The body weight was monitored daily, and the organs were weighed after the euthanasia. Hematological and biochemical determinations were carried out using specific commercial kits and following the protocols provided by the manufacturers. The organs were fixed, sectioned, processed for hematoxylin and eosin staining, and analyzed by light microscopy. Genotoxicity assessment was performed by the alkaline single-cell gel electrophoresis. Livers were processed for ethoxyresorufin-O-deethylase (EROD) and Glutathione S-transferase (GST) assays.

SF1 exhibited low toxicity, as no significant discrepancy was observed in the relative weight of the body organs of mice. Moreover, the daily treatment with SF1 did not alter the number and percentage of red blood cells or hemoglobin concentration in the blood. The treatment with SF1 did not affect the creatinine concentration, but the 25 mg/kg dose reduced the plasma urea level and uric acid, suggesting its use in treating acute renal failure. The parameters analyzed did not present biochemical alterations indicative of liver disease. Regarding serum triglyceride and cholesterol levels, a significant decrease was detected in both parameters in mice treated with SF1. In addition, the histopathological analysis showed that inflammatory focus in the livers seemed more relevant in the control groups than in those treated. There were no significant changes in the renal or splenic tissues of animals treated with SF1. Treatment with SF1 also does not have a genotoxic effect on liver cells.

Treatment with SF1 showed no toxicity in mice at doses equivalent to those recommended for humans, which provides evidence of the safety of the therapeutic use of this subfraction.

The beneficial effects of caloric restriction (CR) and a ketogenic diet (KD) have been previously shown when performed prior to kidney injury. We investigated the effects of CR and KD on fibrosis development after unilateral kidney ischemia/reperfusion (UIR). Post-treatment with CR significantly (p < 0.05) affected blood glucose (2-fold decrease), ketone bodies (3-fold increase), lactate (1.5-fold decrease), and lipids (1.4-fold decrease). In the kidney, CR improved succinate dehydrogenase and malate dehydrogenase activity by 2-fold each, but worsened fibrosis progression. Similar results were shown for the KD, which restored the post-UIR impaired activities of succinate dehydrogenase, malate dehydrogenase, and α-ketoglutarate dehydrogenase (which was decreased 2-fold) but had no effect on fibrosis progression. Thus, our study shows that the use of CR or KD after UIR did not reduce the development of fibrosis, as shown by hydroxyproline content, western-blotting, and RT-PCR, whereas it caused significant metabolic changes in kidney tissue after UIR.

Present study reports an eco and environmental friendly biogenic synthetic protocol for generation of Co₃O₄ nanoparticles (NPs) using aqueous extract of leaves of Ipomoea carnea plant, which act as both reducing and capping agents. Thus produced Co₃O₄ material has been characterized by various techniques like UV–visible absorption spectrum, Fourier transform infrared spectroscopy (FTIR), powder X-ray diffraction (XRD), Scanning electron microscope (SEM) with Energy-dispersive X-ray (EDX),Transmission electron microscope (TEM), (X-Ray Photoelectron Spectroscopy (XPS), Brunauer-Emmett-Teller (BET), particles size distribution analyses. The results revealed formation of well crystalline, mesoporous very small size (6–10 nm diameter), having very narrow size distribution nanoparticles. Phytotoxicity, antifungal and antibacterial activities, lipid test and protein test have been performed using Co₃O₄ NPs. Phytotoxicity results showed the 14.75 ± 0.57 and 9.11 ± 0.86, 7.65 ± 2.110 and 10.6 ± 1.813, and 6.35 ± 3.361 and 12.8 ± 1.720 roots and shoots length of respectively Vignaradiata, Pisumsativum and Cicerarietinum obseved using 20 ppm Co₃O₄ NPs. Antifungal activity was done against A. flavus CR500 while antibacterial activity was performed on Staphylococcus aureus and Shigella flexnerii using MHA media and inhibition zone was recorded till 17.8 mm and 16.7 mm respectively. 96% yield in catalytic oxidation of benzaldehyde into benzoic acid was found using Co₃O₄ NPs as catalyst.

Filter paper provides an excellent matrix for retention of proteins containing a cellulose binding domain. To use this capability for manipulating recombinant fusion proteins, binding and elution parameters were explored and procedures developed for small scale purification, modification and assay. Proteins were tagged with the cellulose binding domain from the Clostridium thermocellum CipB gene via a cleavable linker. Filter paper disks of 6 mm diameter were able to bind up to 80 μg protein although there was a substantial dependence on molecular size. Different means of introducing fusion proteins to the disks allow either binding within 20 min from microliter volumes or slower binding from milliliter volumes. Elution with protease in small volumes yielded greater than 10 μg amounts with concentrations in the 1–2 mg/ml range. To demonstrate their utility, disks were used for small scale protein purification, covalent modification of protein, immunoprecipitation, and in a binding assay. These versatile methods allow parallel processing of multiple samples and may find many uses when only small amounts of protein are needed.

Roadside plant communities are especially exposed to air pollution, including a diverse cocktail of potentially toxic compounds transported by particulates. Some groups of these compounds, heavy metals and polyaromatic hydrocarbons influence plants through reactive oxygen species (ROS) production. Bioindication studies have reported different ecological responses elucidated by these compounds, however, very few comparative studies exist on herbaceous plant species. In our study, the No. 227 OECD GUIDELINE FOR THE TESTING OF CHEMICALS: Terrestrial Plant Test: Vegetative Vigour Test was followed to assess the effect of water soluble components of aerosol on typical members of roadside community. Potential phytotoxicity was assessed based on the following end-points: biomass, chlorophyll a and b, carotenoids and peroxidase (POD) activity. While species-specific sensitivity was experienced of these end-points, considerable ratio (30%) of the test species did not show statistically significant response to the treatment, indicating the relative robustness of roadside communities.