Magnetic pH-responsive nanogels as multifunctional delivery tools for small interfering RNA (siRNA) molecules and iron oxide nanoparticles (IONPs)

We here exploit pH-responsive nanogels as carriers to deliver functional anti-GFP siRNA and superparamagnetic IONPs to HeLa\u2013GFP cells. The siRNA release via pH-mediated endosomal escape is shown. The IONPs act first as magnetofection agents to boost cellular uptake and then as probes to track the release mechanism by electron microscopy

Evaluation of mitochondrial respiratory efficiency during in vitro capacitation of human spermatozoa

The role of mitochondria in sperm motility was the subject of several investigations. However, different views on this topic emerged among scientists. In particular, very little is known on the mechanisms of energy production occurring during human sperm capacitation and related processes. In this study, we have investigated the mitochondrial respiratory efficiency in human sperm samples from normozoospermic subjects before and after swim-up selection and incubation under capacitating condition. Sperm cells, selected by swim-up treatment, were incubated up to 24 h and then demembranated by hypotonic swelling at selected times. The oxygen uptake rate was measured in both basal and swim-up selected samples by a polarographic assay. Mitochondria of swim-up selected cells showed an impressive oxygen consumption rate, which was about 20 times higher than that measured in basal samples. The high mitochondrial respiratory efficiency remained stable up to 24 h after the swimup treatment. The respiration control ratio, the substrate specificity and the inhibitor sensitivity in the swim-up selected samples were similar to those of basal samples thereby suggesting that the physiology of mitochondria was preserved after the swim-up treatment. Furthermore, the remarkably high mitochondrial respiration in swim-up selected samples allowed the oxygraphic analysis of just 200 000 sperm cells. Sperm selection and incubation under capacitating condition are therefore associated with a high activity of the mitochondrial respiratory chain. The sperm oxygen consumption rate could be useful to exclude mitochondria malfunctioning in male infertility

Fucosyltransferase 1 mediates angiogenesis, cell adhesion and rheumatoid arthritis synovial tissue fibroblast proliferation

Abstract Introduction We previously reported that sialyl Lewisy, synthesized by fucosyltransferases, is involved in angiogenesis. Fucosyltransferase 1 (fut1) is an α(1,2)-fucosyltransferase responsible for synthesis of the H blood group and Lewisy antigens. However, the angiogenic involvement of fut 1 in the pathogenesis of rheumatoid arthritis synovial tissue (RA ST) has not been clearly defined. Methods Assay of α(1,2)-linked fucosylated proteins in RA was performed by enzyme-linked lectin assay. Fut1 expression was determined in RA ST samples by immunohistological staining. We performed angiogenic Matrigel assays using a co-culture system of human dermal microvascular endothelial cells (HMVECs) and fut1 small interfering RNA (siRNA) transfected RA synovial fibroblasts. To determine if fut1 played a role in leukocyte retention and cell proliferation in the RA synovium, myeloid THP-1 cell adhesion assays and fut1 siRNA transfected RA synovial fibroblast proliferation assays were performed. Results Total α(1,2)-linked fucosylated proteins in RA ST were significantly higher compared to normal (NL) ST. Fut1 expression on RA ST lining cells positively correlated with ST inflammation. HMVECs from a co-culture system with fut1 siRNA transfected RA synovial fibroblasts exhibited decreased endothelial cell tube formation compared to control siRNA transfected RA synovial fibroblasts. Fut1 siRNA also inhibited myeloid THP-1 adhesion to RA synovial fibroblasts and RA synovial fibroblast proliferation. Conclusions These data show that α(1,2)-linked fucosylated proteins are upregulated in RA ST compared to NL ST. We also show that fut1 in RA synovial fibroblasts is important in angiogenesis, leukocyte-synovial fibroblast adhesion, and synovial fibroblast proliferation, all key processes in the pathogenesis of RA.http://deepblue.lib.umich.edu/bitstream/2027.42/109505/1/13075_2012_Article_4111.pd

Rapporto i.c.a.i. Storia e risultati, di una strategia di ricerca epidemiologica e strumentale

It identifies also a group of vascular surgeons and angiologists, who in the last few years have been undertaking a permanent research program in this area, which is still 'orphan', with respect to the availability of reliable epidemiological data and standardized clinical procedures. Stimulated by the suggestions of an European consensus, the i.c.a.i. group addresses several unsettled issues of CLI: it drew a profile of the different diagnostic and therapeutic interventions used in CLI, in order to assess the consistency of the current procedures with the recommendations of the consensus as well as their practicability. This epidemiological survey served also as a feasibility phase of subsequent intervention studies: it provided information on the prognostic profile of a real population observed in routine conditions of care, thus allowing a careful estimation of the population to be recruited in clinical trials conducted in the same setting. A large scale randomised trial was actually performed, addressing a long unanswered question, it provided evidence of some benefits of the use of prostanoids in CLI. Though small and short-lasting, the advantage is of apparent clinical relevance for a severe clinical condition for which effective treatments were so far unavailable. On its turn, the trial data base (including over 1,500 patients) offers now an epidemiological background suitable to identify the outcome predictive factors in the natural history of CLI, generate further hypothesis related to particular risk subgroups and revise the adequacy of the consensus recommendations