Eco-friendly synthesized silver nanoparticles from endophytic fungus Phyllosticta owaniana: KUMBMDBT-32 and evaluation of biomedical properties.

The primary objective of the current investigation was the biosynthesis of Phy-AgNPs by the endophytic fungus Phyllosticta owaniana (extracted from Abrus precatorius) and the evaluation of the secondary metabolites from the ethyl acetate extract of P. owaniana cultivated by submerged fermentation. Utilizing bioanalytical strategies, Phy-AgNPs were characterized. The UV-visible spectrophotometer analysis revealed an absorption spectrum with a peak at 420 nm, thus validating the Phy-AgNPs synthesis. The FTIR analysis revealed peaks correlating to various potential functional groups, suggesting that Phy-AgNPs have been reduced and capped. SEM-EDAX and HR-TEM analyses demonstrated the spherical shape of Phy-AgNPs, and the 3 keV EDAX analysis confirmed the existence of silver atoms. XRD analyses showed the Phy-AgNPs crystalline structure. The size and the stability of synthesized Phy-AgNPs (65.81 nm) were measured by DLS and Zeta potential studies. While the ethyl acetate extract was analyzed with GC-MS and FTIR for secondary metabolites. The synthesized Phy-AgNPs showed effective antibacterial activity against Pseudomonas aeruginosa (15.1 ± 0.17 mm, 10 mg/mL), while the antifungal activity of Phy-AgNPs inhibited the growth of Candida albicans extremely efficiently (12.16 ± 0.28 mm, 10 mg/mL). Phy-AgNPs were evaluated for a variety of biomedical properties in which they showed significant activity. In a cell viability assay using the MTT assay, Phy-AgNPs exhibited a cytotoxic impact of up to 30.67% and 34.53% when 200 µg/mL were detected. In both in vitro and in vivo anti-inflammatory examinations, nanoparticles (NPs) exhibited a significant anti-inflammatory effect. These findings support the pharmaceutical and biomedical properties of the synthesized Phy-AgNPs.

Estimation of serum hyaluronidase activity overcoming the turbidity interference.

The assay of mammalian hyaluronidases (HAases) is important in understanding the role of the hyaluronan-hyaluronidase (HA-HAase) system in various pathophysiological processes. Despite several quantitative assay method options, the Morgan-Elson colorimetric method modified by Reissig et al is considered the best for determining the activity in clinical samples. However, the sensitivity of the method was greatly limited by presence of protein above 400 microg due to turbidity interference that led to chromogen quenching. Therefore, an effort has been made to reinvestigate the Reissig et al method. In the reinvestigated method, a standardized optimal 0.32 M potassium tetraborate (PTB) was used against 0.13 M (native) to overcome the turbidity interference. The estimated mean OD at 585 nm of serum for native method was 0.043 (95% CI: 0.040 to 0.045), while that for the re-investigated method was 0.138 (95% CI: 0.133 to 0.143, p < 0.0001). The mean OD at 585 nm of serum of native method was significantly lower than that of re-investigated method (p < 0.05) at all protein levels. This was also true for estimated mean OD at 585 nm of plasma. The mean intrasample CVs for native and re-investigated methods were 0.9% and 0.5%, respectively, for normal serum. Furthermore, the repeatability coefficient of normal serum for native was 0.003 IU, while re-investigated method experienced that of 0.002 IU.

The anti-ophidian properties of Anacardium occidentale bark extract.

Snakebites in rural areas of tropical and subtropical regions are commonly treated with medicinal plants. In this report, we have studied the ability of Anacardium occidentale bark extract to neutralize enzymatic as well as pharmacological effects induced by Vipera russelii venom. The extract neutralized the viper venom hydrolytic enzymes such as phospholipase, protease, and hyaluronidase in a dose dependent manner. These enzymes are responsible for both local effects of envenomation such as local tissue damage, inflammation and myonecrosis, and systemic effects including dysfunction of vital organs and alteration in the coagulation components. In addition, extract neutralized the pharmacological effects such as edema, hemorrhage, and myotoxic effects including lethality, induced by venom. Since, it inhibits both hydrolytic enzymes and pharmacological effects; it may be used as an alternative treatment to serum therapy and, in addition, as a rich source of potential inhibitors of hydrolytic enzymes involved in several physiopathological diseases.

Purification and properties of hyaluronidase from Hippasa partita (funnel web spider) venom gland extract.

Spider venom is a complex mixture of protein and peptide toxins. Hyaluronidase a 'spreading factor' has not been studied extensively in spider venom. In this paper, we describe the purification and characterization of a hyaluronidase from Hippasa partita venom gland extract. Hyaluronidase (HPHyal) has been purified by the successive chromatography on a Sephadex G-100 and on CM-Sephadex C-25 columns. HPHyal has been purified to an extent of about approximately 20-folds. The molecular mass was found to be 42.26 kDa by matrix-assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometry. HPHyal was optimally active at pH 5.8 at 37 degrees C and in the presence of 300 mM NaCl in the reaction mixture. HPHyal showed absolute specificity for hyaluronan and belongs to neutral active group of enzymes. HPHyal revealed single-precipitin line, while venom gland extract revealed multiple bands in Western blotting with the antiserum prepared against venom gland extract. HPHyal indirectly potentiates the myotoxicity of VRV-PL-VIII myotoxin and also the hemorrhagic potency of hemorrhagic complex-I. Cations, Na(+) and K(+) enhanced the activity and chloride ions do not have any effect while, divalent cations, inhibited the enzyme activity.