Analysis of cerebrospinal fluid (CSF) facilitates the understanding of brain-specific molecular changes that may associate with disease progression. Proximity extension assays (PEA) have been deployed in several CSF studies, however the evaluation of the assay and impact of freeze-thaw cycles on the protein signal has not been documented. We sought to (1) assess the technical performance of the PEA platform and (2) evaluate the effect of freeze-thaw cycles on the detectability of analytes on the PEA platform. We have evaluated the PEA with Next Generation Sequencing (NGS) readout assay and report on the detectability and coefficient of variation observed in CSF samples. We have also evaluated proteomic signals with a minimum of 3 and a maximum of 9 freeze thaw cycles and detected very minimal change in signal with increasing cycle number. Our study is the first to evaluate the performance of PEA using NGS readout platform with CSF samples. We report lower protein detection rates and higher variability in the expansion panels compared to the original 4 panels, with acceptable variation above detectability threshold. In addition, our work demonstrates that the proteomic signal is robust and continues to be stable across multiple freeze thaw cycles. This is highly impactful to the processing and analysis of clinical samples and facilitates the investigation of samples with variable pre-analytical conditions.
Diosmin is a flavonoid glycoside renowned for its potent antioxidant, anti-hypertensive, anti-inflammatory, and anti-cancer properties. However, its natural abundance is limited, necessitating efficient production strategies. The objective of the present study was to evaluate the efficacy of Piriformospora indica mycelium extract as a biotic elicitor to enhance the biosynthesis of diosmin and other polyphenolic compounds in Scrophularia striata cell suspension cultures, comparing the efficiency of shake-flask and bioreactor systems. P. indica mycelium extract (250 mg l− 1) was applied at concentrations of 0, 0.5, 1, and 2
Health concerns about bisphenol A have prompted its replacement with analogues, bisphenol F (BPF) and bisphenol S (BPS). Regrettably, there is paucity of data on potential health hazards of these analogues while research on their removal from aqueous media is still emerging. In this study, effects of BPF and BPS exposure at sublethal concentration and bioremediation treatment were investigated on biochemical, hematological and histological parameters of selected organs (gills, liver, kidney, muscle and brain) of juvenile African catfish. 10
Vector-borne diseases transmitted by Aedes mosquitoes remain a major global public health concern. With the growing incidence of insecticide resistance and the potential environmental hazards posed by chemical insecticides, the development of effective green and eco-friendly alternatives to chemical products for the control of mosquito-borne diseases is of paramount importance. The objective of this research work was to investigate the mosquitocidal activities of Dracaena angolesnsis leaves and associated expression of resistance-related genes and their toxicity against Aedes aegypti mosquitoes. Phytochemical screening revealed the presence of several bioactive compounds, including flavonoids, phenols, terpenoids, sterols, tannins, and alkaloids, which are known for their insecticidal potential. Larvicidal, pupicidal, and adulticidal bioassays demonstrated significant mosquito mortality across different solvent extracts, with aqueous and methanol extracts exhibiting the highest efficacy. The lowest larval LC₅₀ value was observed for the aqueous extract (12.97 ppm), while strong pupicidal and adulticidal activities were also recorded. RT-qPCR analysis of the voltage-gated sodium channel (kdr) transcript revealed marked upregulation in treated larvae, with expression levels increasing from 90.65-fold to 10,524-fold relative to untreated control larvae from the same laboratory-maintained field strain. Complete adult mortality (100
Streptococcus suis serotype 31 (SS31) is a clinically important swine pathogen for which inactivated vaccine development requires a scalable cultivation process and evidence that the resulting bacterin has protective activity. However, conventional culture processes often yield insufficient biomass, whereas aggressive high-density fermentation may perturb the physiological features that determine vaccine quality. Herein, we demonstrate a Response Surface Methodology (RSM)-guided fermentation-to-vaccine strategy that optimizes the SS31 culture microenvironment and evaluates the protective activity of a bacterin produced under the optimized conditions. Single-factor screening and Box-Behnken modeling identified glucose, organic nitrogen and newborn bovine serum as dominant growth variables, leading to an optimized formulation containing 2.19 g/L glucose, 43.8 g/L organic nitrogen and 5.89
