Journal of Bioanalysis & Biomedicine

Many cell types release exosomes and microvesicles (EMVs) with a size range from 0.05-5 μm, harbouring receptors, bioactive and signaling proteins, molecular mediators and nucleic acids for cell-to-cell communication. Microvesicles bud directly from the plasma membrane, in contrast to exosomes which are derived from multivesicular bodies within the cell. Increased levels of EMVs have been observed in plasma, urine and other body fluids in patients suffering from a wide range of common complex diseases, including vascular, metabolic, lung, autoimmune and neurodegenerative diseases, chronic inflammation and cancer. EMVs may affect target cells directly by surface-bound ligands, transferred surface receptors and membrane-associated enzymes, such as glycosylphosphatidylinositolanchored proteins, or delivered cytoplasmic or membrane-associated constituents, such as cytosolic proteins, micro/mRNAs, bioactive lipids and even mitochondria. The use of EMVs as diagnostic markers for the prediction, diagnosis, therapy monitoring and prognosis of complex diseases is becoming increasingly attractive. Novel technologies for analysis of the size, density and molecular composition of EMVs are currently emerging together with methods for their improved isolation and purification out of heterogenous vesicle populations. In addition, the recent revolution in mass-spectroscopy, (micro-) flow cytometry, atomic force microscopy, nanoparticle tracking and biosensing will considerably facilitate the quantitative and qualitative analysis of all the constituents assembled in EMVs. Technologies will be preferred that provide signatures specific for EMV subsets rather than a single or a few parameter(s) averaged for the total EMV population. Those EMV signatures have to be correlated to specific disease states along cross-sectional and longitudinal clinical studies. Moreover, it has to be tested which signatures and molecular components, i.e. EMV subspecies, are most informative to obtain actionable disease information. Ultimately, the reliable, rapid and low-cost analysis of EMVs will support systems biology-based approaches for the diagnosis and therapy of complex diseases and supplements the analytical power of conventional biomarkers.

An aim of the study is development and validation of a method for the simultaneous estimation of raloxifene (RAL) and its two active metabolites, raloxifene-4-glucuronide (R4G) and raloxifene-6-glucuronide (R6G) in human plasma samples using raloxifene-D4 as an Internal Standard. Sample preparation was performed by solid phase extraction (SPE) and was followed by separation of the analytes on a UPLC system with a linear gradient and a mobile phase consisting of acetonitrile and ammonium formate. Detection was achieved by tandem mass spectrometry operated in the electrospray positive ion mode. The method had a short sample preparation time, as well as a chromatographic run time of just 4.2 min, the shortest so far reported for RAL, R4G and R6G determination. It was validated and fulfilled all preset criteria for sensitivity, specificity, linearity, within inter- and intra-accuracy and precision, stability studies for all molecules. The method was linear in the concentration range of 0.040 to 1.5 ng/mL, 0.6 to 50.0 ng/mL and 0.6 to 50.0 ng/mL for RAL, R4G and R6G, respectively. The proposed method could be applied to the rapid and reliable simultaneous determination of RAL, R4G and R6G in a bioequivalence study.

Background: Ceiba pentandra seeds belonging to order Malvalea and the family Malvaceae, commonly known Kapok were evaluated for their phytochemical ingredients and antioxidant activity.

Materials and methods: Soxhlet extraction method was used for the extraction of oil, phytochemicals and antioxidant activities of Ceiba pentandra seed oil was estimated by well-known methods.

Results: The seed oil was extracted using soxhlet extraction method with analytical grade hexane as refluxing solvent. The oil was thick yellowish in colour having pungent odour with a yield of 40%. Phytochemical constituents and non enzymatic antioxidants showed enhancement in increasing concentration from 25 mg/mL to 100 mg/mL. At 100 mg/mL recorded phytochemical components phenols, flavonoids, alkaloids and tannins were 305 μg gallic acid equivalents g^-1, 3207 μg quercetin equivalents g^-1, 0.34 μg boldine equivalents g^-1 and 2041 μg tannic acid equivalents g^-1 respectively. The seed oil exhibited striking Diphenyl picryl hydrazyl radical scavenging Assay (DPPH), Ferric reducing or Antioxidant power assay (FRAP), reducing power assay and Hydroxyl radical scavenging activity. The measured DPPH activity at 100 mg/mL was 47.56% of inhibition per 50 μL of oil as compared to 45 and 76% of inhibition per 1 mg/mL of Rutin and BHT as positive controls. Observed FRAP ability of Ceiba pentandra seed oil was 309 FRAP units at 100 mg/mL concentration, reducing activity of Ceiba pentandra seed oil was observed at 100 mg/ mL as 20.52 μg of ascorbic acid equivalents per mL of oil. Hydroxy radical scavenging activity of Ceiba pentandra seed oil observed as 39.69% inhibition/0.1 of oil at 100 mg/mL, as compared to positive controls, Ascorbic acid and BHT at 1 mg/mL with percentage of inhibition 75 and 75.6 respectively.

Conclusion: The importance of the phytochemical constituents and non-enzymatic activities of Ceiba pentandra seed oil in the maintenance of health is strengthened as trend of the future is moving towards using seed oil as medicine in the management of various chronic diseases.

Analysis of pesticides in house dust, as an index of environmental pesticide exposure, is useful in the evaluation of pesticide effects in children. This study compares the prevalence and concentrations of pesticides (propoxur, transfluthrin, bioallethrin, cyfluthrin, and cypermethrin) in house dust collected by the HVS3 vacuum and by sweeping using the house broom. The pesticides were extracted from the dust samples by solid phase extraction and analyzed by gas chromatography/mass spectroscopy. There were significant correlations between the pesticides found in the swept and vacuumed samples (kappa=0.28 to 0.48, rho=0.31 to 0.55). Significantly higher prevalence and concentration of propoxur and higher concentrations of pyrethroids were found in the swept compared to vacuum dust samples. We conclude that ongoing exposure of children to pesticides can be monitored by the analysis of house dust collected by sweeping. Sweeping offers an excellent alternative for house dust collection in areas where vacuum collection is not feasible.