LeukoCatch, a quick and efficient tool for the preparation of leukocyte extracts from blood

<p>Abstract</p> <p>Background</p> <p>Whole-protein extracts from peripheral blood leukocytes are ideal for basic and clinical research. However, lack of a simple preparation technique has limited the use of such extracts. The aim of this study is to develop a simple and easy system that can selectively obtain leukocyte extracts without hemoglobin.</p> <p>Methods</p> <p>A filter that captures the leukocytes but not RBCs was set at the bottom of a 10-mL medical syringe by sandwiching it between plastic stoppers. The capturing efficiency of leukocytes with this tool, called LeukoCatch, was examined using human macrophage cells (MONO-MAC-6). The abilities of LeukoCatch system to capture the leukocyte proteins and to remove the hemoglobin from RBCs were tested by western blot analysis using human blood samples.</p> <p>Results</p> <p>This study presents the development of LeukoCatch, a novel tool that allows the preparation of leukocyte extracts from blood samples within 3 min without centrifugation. Tissue-cultured human macrophage cells were tested to determine the optimal filter numbers and pass-through frequencies of LeukoCatch, which was then applied to 2-mL blood samples. Samples were passed 2~5 times through a LeukoCatch equipped with 5 filters, washed twice with phosphate-buffered saline for red cell removal, and leukocyte proteins were extracted with 0.5 mL of elution buffer. Western blot analysis of the purified extract indicated that more than 90% of hemoglobin was removed by the LeukoCatch and that the protein recovery rate of leukocytes was at least 4 times better than that of the conventional centrifugation method.</p> <p>Conclusion</p> <p>We conclude that LeukoCatch is useful not only for diagnosis at the bedside but also for basic research using blood samples or tissue culture cells.</p

Neuropsychological function is related to irritable bowel syndrome in women with premenstrual syndrome and dysmenorrhea

Background There is increasing evidence demonstrating the co-occurrence of primary dysmenorrhea (PD), premenstrual syndrome (PMS), and irritable bowel syndrome (IBS) in women. This study aimed to investigate whether women who have symptoms of IBS in addition to PD and PMS also report more severe or frequent menstruation-associated symptoms and psychological complications compared to women with PD and PMS alone. Methods The study group included 182 female University students aged 18–25 years. IBS was diagnosed using the Rome III criteria. The severity of PMS and PD was determined using a 10-point visual analog scale and PSST (Premenstrual Syndrome Screening Tool), respectively. Neuropsychological functions including cognitive function, depression score, anxiety score, stress, insomnia, daytime sleepiness, quality of life and personality were assessed using standard questionnaires. Results Of the 182 young females, 31 (17.0%) had IBS. Average days of bleeding during the menstrual cycle and mean pain severity on the PSST scale were significantly greater in the group with IBS compared to the non-IBS group (p < 0.01). The non-IBS individuals scored more favorably than the women with IBS with respect to severity of depression, insomnia, daytime sleepiness (p < 0.05). The PSST scores were significantly correlated with scores for depression (r = 0.29; p < 0.001), anxiety (r = 0.28; p < 0.001), stress (r = 0.32; p < 0.001), insomnia (r = 0.34; p < 0.001) and daytime sleepiness (r = 0.31; p < 0.001); while, they were negatively correlated with cognitive abilities (r = − 0.20; p = 0.006) and quality of life (r = − 0.42; p < 0.001). Linear regression analysis showed that the PSST scores were possibly significant factors in determining the scores for depression, anxiety, stress, quality of life, insomnia and daytime sleepiness (p < 0.05). Conclusion IBS is related to psychological comorbidities, in particular depression, sleep problems and menstrual-associated disorders. IBS may exacerbate the features of PMS which should be taken into account in the management of PMS

Particle affinity and bioavailability of PAHs associated with coal tar pitch

This project has investigated the particle affinity and bioavailability of coal tar pitch related PAHs from sediments outside several Nordic aluminium smelters using the Søderberg technology. Passive samplers (POM) have been used for assessing water-sediment partitioning for PAHs and actual bioaccumulation has been addressed in an experimental setup.The results showed that the PAHs associated with the sediments in the vicinity of the smelters were stronger (a median factor of at least a magnitude) adsorbed/absorbed to the particles than free energy relationship implies. This further implies that the bioavailable fraction is correspondingly lower, and one would expect lower bioaccumulated concentrations. The accumulated concentrations measured in Nereis diversicolor and Hinia reticulata were in fact very similar to biota concentrations expected based on the POM-deduced sediment-water partitioning coefficients. Thus, the measured biota to sediment accumulation factors (BSAFs) agreed also very well with those expected from the POM-deduced sediment-water partitioning coefficients. On the other hand, this good correspondence was not observed for the third species, the bivalve Nuculoma tenuis. There were however logistical intractabilities connected to this species biology and size that render it probable that particulate sedimentary matter contaminated the Nuculoma tissues analyses. The uncertainties associated with Nuculoma tenuis render the interpretations somewhat inconclusive for this species, and further investigations would be necessary to eliminate the uncertainties

Biotilgjengelighet av miljøgifter i sedimenter fra Bredalsholmen, samt toksisitet av sedimentene for algen Skeletonema costatum

NIVA har på oppdrag fra Kristiansand kommune undersøkt biotilgjengelighet og toksisitet av miljøgifter i sedimenter nær Bredalsholmen. Blåskjell fra området har også vært analysert. Blåskjellanalysene viser en viss PAH-påvirkning på området, sannsynligvis fra lokale tilførsler (båthavn) og transport fra andre områder i indre Kristiansandsfjorden. Det var ikke indikasjoner på aktive tilførsler av PCB til de øvre vannlag i området. Forhøyede HCB-verdier kan skyldes at sedimentene i indre Kristiansandsfjorden generelt er påvirket av HCB. Sedimentene fra området rundt Bredalsholmen var sterkt forurenset med PAH og man kunne observere akkumulering i organismer eksponert for disse. Imidlertid var ikke PAH-konsentrasjonene i organismene veldig mye høyere enn i kontrollgruppen. Sedimentene var kun moderat forurenset med PCB, men akkumulering i organismer ble vist. Resultatene viste indikasjoner på at PAH var sterkere partikkelbundet, enn PCB. Toksisitetstester av porevann med kiselalgen Skeletonema costatum viste at porevannet hadde veksthemmende effekter på alger. Toksisiteten var imidlertid lavere enn grenseverdien som er foreslått i SFTs veileder for risikovurdering av forurensede sedimenter på alle stasjonene som ble undersøkt. På den ene stasjonen var toksisiteten nær grensen